PATENT DOCUMENT

Abstract:
This application pertains to construction of pooled biological material such as DNA, RNA, proteins and the like that are able to be screened by a wide variety of methods such as PCR (Polymerase Chain Reaction), DNA/DNA hybridization, DNA/RNA hybridization, RNA/RNA hybridization, single strand DNA probing, protein/protein hybridization and a wide variety of additional methods. Our new method for construction of pools and superpools for screening differs in that the complete set is systematically divided into a variety of smaller subsets which are then re-pooled to make the final screening pools. This pooled material can be from individual samples or a population of samples. In order to reduce the analysis time, materials and expense, the pooling of high resolution small pools in a matrix allows for a lower number of user experiments to have higher resolution (as if the researcher had analyzed the complete set of small pools).

Full Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     This application claims the benefit of U.S. Provisional Patent Application Ser. No. 60/467,912, filed May 5, 2003 and entitled “Pool and Superpool matrix provisional application” now abandoned, which is herein incorporated by reference in its entirety for all purposes. 
    
    
     STATEMENT REGARDING FEDERALLY SPONSERED RESEARCH OR DEVELOPMENT 
     Not Applicable 
     REFERENCE TO SEQUENCE LISTING, A TABLE, OR A COMPUTER PROGRAM LISTING COMPACT DISK APPENDIX 
     Not Applicable 
     Field of Search: 341/., 435/., 435/ . . . , 435/DIG21, 435/DIG51, 536/23.1, 700/., 707/101 
     References Cited [Referenced By] 
     U.S. Patent Documents 
     5,780,222 
     6,126074 
     6,477,699 
     6,582,923 
     6,607,888 
     6,665,829 
     6,706,867 
     6,727,068 
     6,727,071 
     Other References 
     Ausubel et al., “Short Protocols in Molecular Biology”, Wiley and Sons, New York. 
     Sambrook et al., “Molecular Cloning, A Laboratory Manual”, Cold Springs Harbor Press, New York. 
     Torney et al., “Pooling of a Total Genomic BAC Library”, US DOE Contract W-7405-ENG-36. 
     Borm, T. J. A., BACBank on the Internet http://137.224.204.155/bacbank (also copied in provisional application 60/467,912). 
     BACKGROUND OF THE INVENTION 
     This application pertains to construction of pooled biological material such as DNA, RNA, proteins and the like that are able to be screened by a wide variety of methods such as PCR (Polymerase Chain Reaction), DNA/DNA hybridization, DNA/RNA hybridization, RNA/RNA hybridization, single strand DNA probing, protein/protein hybridization and a wide variety of additional methods. References describing many of these methods include “Ausubel et.al. Short Protocols in Molecular Biology, Wiley and Sons, New York” and “Sambrook et.al, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, New York” as well as numerous others and are hereby included by reference. Also included by reference are U.S. Pat. No. 5,780,222 (Method of PCR Testing of Pooled Blood Samples) and its references cited. Also included are U.S. Pat. Nos. 6,126,074 and 6,477,669 and their references including the references pertaining to Veterbi, Reed-Solomon and other Error Correction and Data Compression Coding schemes. This pooling method will allow the incorporation of ‘loss-less information compression and error correction’ or other ‘current art’ error correction strategies to improve the robustness of identification with significantly reduced numbers of samples to be processed by the end user. By having the samples pooled again after collection, it is possible to drastically reduce the manipulations required by the end user while still keeping very fine detail in the identification of the individual samples or populations that were originally pooled. These error-correction methods are well known in the computer data transmission field, but have not been used in the pooling of biological or chemical samples. The use of these methods will allow a large reduction in the number of experiments required to identify the specific biological sample or population containing a region of interest. 
     This pooled material can be from individuals or a population. In order to reduce the analysis time, materials and expense, the pooling of small high resolution pools in a matrix allows for a lower number of samples to be analyzed. The resulting high resolution data obtained from screening these matrix pools are equivalent to the data obtained if the researcher had analyzed the complete set of small pools (much more expensive, time consuming and difficult). This method also gives the added advantage of having two positive signals needed for identification. This reduces the problems associated with a false positive when only one signal is obtained for identification (as in the Current Art). 
     This matrix pooling can be just in one superpool. Alternatively, it can be a matrix of a variety of different superpools and/or across a variety of different types of pools to allow the screening of the complete library with just one round of experiments. To do this, each small pool would be added to between 6 and 20 of the collection of re-pooled intermediate or final pools. Then with the total number of pools of between 40 and 100, the complete library (or any set of biological samples) could be screened with high confidence and the ability to resolve multiple hits. If the library had a large redundancy of signal, the total number of pools could be increased to maintain accurate resolving power of the matrix method. The incorporation of positive controls in a matrix pattern can be used for quality assurance and for assisting in deconvolution if desired. 
     The current state of the art in pooling of biological materials such as Bacterial Artificial Chromosome (BAC) genomic DNA libraries (and other biological or chemical libraries like cDNA libraries, protein libraries, RNA libraries, DNA libraries cellular metabolic libraries and chemical libraries) for screening consist of the collection of all of the indexed microtiter plates containing the BAC library and then forming these plates into a large cube. These indexed plates are generally 96, 384, 864 well or sometimes even 1536 well microtiter plates. This large cube is then transected by a number of different planes (usually 4 to 8) which produce a large number of pools from each plane. This collection of all of the pools from all of the planes are then screened to identify the clones of interest. This scheme is the current state-of-the-art and can identify multiple clone hits with some degree of reliability to identify multiple targets (i.e. BAC clones) at a specific coordinate. According to Klein et al., their scheme with 6 planes in a collection of 24,576 BAC&#39;s could detect between 2 and 6 BAC&#39;s and over 90% could be reliably assigned to a specific coordinate with 184 screening pools (that is 184 user experiments are required). 
     Prior art as disclosed in S. Asakawa et al. Human BAC Library: Construction and Rapid Screening, Gene Vol. 191, pp. 69-79 (1979), may disclose some of the initial steps that are similar to the present invention in the Methods I section on page 72 but requires pooling clones before growth and requires construction of each screening pool directly from the pooled clones after growth. 
     The reason for the present invention accuracy, efficiency and reduced cost is that the present invention requires at least one additional step of repooling the intermediate subpooled genomic DNA clone DNA into a final screening pool, where the individual genomic DNA clone is in between three to ten unique final screening pools or between at least 4 unique Matrix Pools and no more than 8 unique Matrix Pools. 
     If the BAC Library is from an organism with a genome larger than 1,000 Mb, the researcher may find that there are very few ambiguous hits in the plate, row, column and diagonal (PRCD) plate. The Plate, Row, and Column pools correctly identify the clone of interest without the need for the Diagonal Pools. If the Diagonal Pools are only screened to solve the infrequent ambiguity, there would be a reduction in the number of PCR experiments. 
     A Bac-Bank is a way of storing fragments of DNA, together constituting the whole genome of an organism. The DNA of an organism is (semi) randomly cut in pieces, and these fragments are inserted into bacteria, which are then plated out so that a single colony grows from a single modified bacterium. Only modified bacteria are allowed to grow by using a bacterium that is potentially resistant to a certain antibiotic, and whose resistance is “switched on” by the presence of a foreign DNA fragment (insert), and by using a growth medium containing the antibiotic. The resulting (potentially) unique colonies of bacteria are then picked up individually and transferred to the wells of 384-well plates, and the resulting stack of plates holding a large number of unique bacteria, ideally containing the whole genome of the original organism, is known as a “Bac-Bank”. It serves as a research database of the genome of the original organism. This database can be searched for fragments of DNA using PCR techniques. 
     Pooling is a method that allows one to quickly and economically search a Bac-Bank for the presence of certain DNA fragments. A Bac-Bank normally contains a large number of clones (˜100,000), and testing all these clones individually for the presence of a fragment of DNA occurring only a few times (typically less than 100 times) in the original organism&#39;s genome is prohibitively expensive and laborious. When pooling is used the DNA of several clones is gathered into a much lower number of wells (pools), every well containing DNA from several clones and every clone&#39;s DNA being present in multiple wells. The distribution pattern (“pooling method”, “pooling strategy”,“rule-set”) is designed in such way that when using PCR reactions to screen the pools a pattern (of PCR reaction results) emerges that is (hopefully) unique to the clone(s) having the required properties. A simple example: take a 384-well plate having 16 rows of 24 columns; imagine pooling all wells horizontally and vertically, resulting in 16 row-pools and 24 column pools. If a single clone in this plate has a certain property, only the column-pool and the row-pool that particular clone is in will display a positive reaction when screened; the other 38 pools will be negative. Using only 40 PCR reaction it is therefore possible to pinpoint the positive clone in this 384-well plate; almost a tenfold reduction in labour and cost. As long as there are relatively few individuals with a certain property there is no problem; for properties that are shared among many individuals all pooling methods break down (yield incorrect results, either false-positives or (worse) false-negatives), and when this happens one has to resort to screening the clones individually. 
     Most often the individual clones in Bac-Bank are identified/labeled according to some hierarchical structure dictated by the physical properties of the Bac-Bank. The number of dimensions of a Bac-Bank is then related to the hierarchical structure of the storage format. 
     An example: The clones of a Bac-Bank are individually stored in wells on a plate. The wells are arranged in a rectangular pattern of rows and columns. If this plate constitutes the whole Bac-Bank this Bac-Bank can be viewed as one-dimensional if all the wells on the plate have consecutive numbers from left to right and top to bottom. One single parameter (well number) suffices to address every individual clone/well on the plate, and therefore the Bac-Bank is one-dimensional. A more natural approach in this example would be to address each well by its column and row numbers; then we would need two parameters to address an individual well, and therefore the same Bac-Bank can be two-dimensional as well. 
     For a larger Bac-Bank one plate would not suffice, and we could give each plate a separate ID code. This would add one coordinate to the number of coordinates required to address each individual well, and therefore there is one more dimension this case than there would before a single plate. Another approach is to store the well-plates in boxes of (for example) 10; each plate itself would then have two parameters (coordinates) for an address: the box number and (within that box) the plate number. 
     All these items are a matter or choice, and therefore the number of dimensions of a Bac-Bank is a choice as well; it is even possible to use several different addressing schemes without imposing any structure upon this number/code, but one may also choose to address an individual well as “[C,  23 ,  4 , A,  6 ]”, when the clone is located in fridge C, box 23, plate 4, column A, and row 6. 
     Having said this, it is important to note that it is most convenient to have some sort of logical structure related to the physical location of a clone; this helps you find individual clones faster, and most often there is also a relation between this logical/physical organization and the way the Bac-Bank is pooled. In all other examples we will assume that the Bac-Bank consists of 300 plates of 24×16 wells, and that the Bac-Bank is three-dimensional. 
     It will be readily apparent that there are a number of important claims that will arise from this disclosure, including but not limited to:
     1. Higher resolution deconvolution of complex data without as many analysis reactions.   2. Analyzing a two, three or more-dimensional matrix of pools allows significant reduction in analysis reactions while retaining a high degree of specificity.   3. The incorporation of loss-less compression and error-correction into the pooling strategy allows improved robustness of analysis and identification if individuals from the pools with increased effectiveness while reducing the numbers of analysis.   4. Significantly reducing the number of analysis reactions required from other, less sophisticated pooling systems if a matrix re-pooling design is utilized.   5. As the analytical methods improve, the ability of re-pooling pools (that currently are at the limits of detection) is another significant improvement and advantage.   

     BRIEF SUMMARY OF THE INVENTION 
     Our new method differs in that the complete set is systematically divided into smaller subsets which are then re-pooled to make the final screening pools. This pooled material can be from individual samples or a population of samples. In order to reduce the analysis time, materials and expense, the pooling of high resolution small pools in a matrix allows for a lower number of user experiments to have higher resolution (as if the researcher had analyzed the complete set of small pools). One of the preferred embodiments describes a two step method that first screens for which superpool. Then that specific superpool&#39;s pools have been re-pooled into matrix pools (which are 36 matrix pools instead of 76 pools). The matrix pools are screened in this method also gives the added advantage of having two or more positive signals needed for identification (as shown in our provisional application). This reduces the current state-of-the-art problems associated with a false positive and/or false negative experimental result when only one signal is obtained for identification. 
     The Round I PCR is performed on all of the subpools “containing all BAC clones in the Library”. Each Superpool contains 4,608 individual BAC clones. The results from Round I of PCR will identify which Superpool BAC clone(s) with the sequence of interest (there may be more than one Superpool identified). The researcher may chose to pursue one or more positive hits from the Round I PCR. 
     The Round II PCR will be performed on the Matrix Pools for the specific Superpool identified in Round I PCR. Round II PCR requires 36 PCR experiments plus controls (for each positive hit pursued from Round I PCR). The results from Round II PCR should allow the researcher to identify the plate and well position for several positive hits and to rule out many potential false positives (in the particular Superpool(s) being pursued). In comparison, using a traditional plate/row/column/diagonal strategy, Round II PCR screening of PRCD pools requires 76 PCR reactions plus controls. The Matrix system reduces the PCR experiments by 50%. 
     The Matrix Pools are PRCD pools combined so that EACH of these PRCD pools is contained in TWO unique Matrix Pools. There are a total of 36 Matrix Pools for each Superpool. Eight Matrix Plate Pools (MPP), eight Matrix Row Pools (MRP), 10 Matrix Column Pools (MCP) and 10 Matrix Diagonal Pools (MDP). There are at most 1,152 individual BAC clones inside each Matrix Pool well. 
     The matrix pooling can be just in one superpool (as we have shown in the detailed description of our matrix manual in our provisional application). Alternately, it can be a matrix of a variety of different superpools and/or across a variety of different types of pools to allow the screening of the complete library with just one round of experiments. To do this, each small pool would be combined with any number (generally between six and many thousands depending on the sensitivity/robustness of the users experimental screening strategy) of final collection pools (which are re-pooled intermediate pools). For this example we&#39;ll use the range of 6 and 20 collection pools (fully compatible with a PCR based screening technology). Then with the total number of pools of between 40 and 180, and more preferably between 80 and 96, the complete library could be screened with high confidence and the ability to resolve multiple samples in the library containing an identical region of interest. If the library had a large redundancy of signal, the total number of pools could be increased to maintain accurate resolving power of the matrix methodology. The incorporation of positive controls in a matrix pattern can be used to for quality assurance and for assisting in deconvolution if desired. 
    
    
     
       BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS 
         FIG. 1  is an entire BAC Library comprised of BAC clones in individual wells of 120, 384-well plates, designated as Superpools 1-10 (SP1-10). 
         FIG. 2  is the Library Code Superpool Collection plate copy #1 having 96 wells. 
         FIG. 3A  is the SP 1  with 12 plates having a clone address in plate #8. 
         FIG. 3B  is the SP 1  with 16 rows having a clone address in row #4. 
         FIG. 3C  is the SP 1  with 24 columns having a clone address in column #16. 
         FIG. 3D  is the SP 1  with 24 diagonals having a clone address in diagonal #6 as specified in table 4. 
         FIG. 4  is the small high resolution pools of the plate, row, column, and diagonal samples of DNA, cDNA or proteins with a positive and negative control in wells E 2  and F 2  respectively. 
         FIG. 5  is the further re-pooling a subset of the wells of  FIG. 4  onto the Matrix Pool Plate or specific repooling designs in a Plate, Row, Column, and Diagonal matrix loading pattern. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       FIGS. 1-5  are a graphical representation of an embodiment of the present invention. Example 2, Tables 6 and 7 are alternate embodiments of  FIG. 3D  and  FIG. 4  respectively. Tables 8-11 represent additional alternative embodiments of specific repooling designs as depicted in  FIG. 5 . And Tables 12-16 are data from additional embodiments of the design techniques.  FIG. 1  represents  10  Superpools of the entire  10  BAC Library containing 120, 384 well-plates  2  stacked on top of each other in 10 sets of 12 plates. A wide, almost limitless set of indexed microtiter plates may be used for the plates  2 . 
     After compiling the Entire BAC Library the researcher receives two identical Superpool Collection Plates that are then used for Round I PCR. 
     Specifically,  FIG. 1  is a combined stack of 10 Superpools (SP 1 -SP 10 ). Each Superpool has a stack of 12 plates  2  stacked upon each other. The plates could be any multi-well unit that can be arranged into a hierarchical structure. Claimed herein are 96, 384, 864 and 1536-well units. 
     In  FIG. 2  a 96-well Superpool Plate  20  comes with a positive A 1  and a negative control B 1  4 and 6 respectively and a sample from each Superpool  8  and each Superpool plate provides the template for at least 800 PCR experiments. 
     After receiving Round I PCR gel electrophoresis results, the researcher determines which Superpool to screen for Round II PCR. 
     In  FIG. 3  each Superpool SP- 1  to SP- 10  is then separated into pools of plates., rows, columns, and diagonals, which are all based on the hierarchical structure for the clone of interest to allow the researcher to find the specific coordinate or unique address of the well position with the clone of interest. At least three of these four hierarchical structures (plate, row, column, and diagonals) must be used or any combination of three of the four hierarchical structures, to insure or guarantee finding the specific coordinate well position with the clone of interest through iteration or redundancy (eg.  FIG. 3D  diagonal pool, plus  FIG. 3A  plate pool, plus  FIG. 3C  column pool). Again  FIGS. 3A-3D  represents the primary embodiment of a Superpool with a stack of 12 plates with 384 wells. The essence of  FIG. 3  (A-D) is to use four different search patterns to find the precise well having the clone of interest.  FIG.3A  identifies the plate of interest in Superpool 1,  30  eg.P- 8 .  FIG. 3B  identifies the row of interest in SP 1 ,  40 , eg. R- 4 .  FIG. 3C  identifies the column of interest in SP 1 ,  50 , eg. C- 16 .  FIG. 3D  identifies the diagonal of interest in SP 1 ,  60 , eg. D- 6 . 
     All of the DNA or protein samples in a Superpool from the plates  30 , rows  40 , columns  50 , and diagonals  60  are sequentially pooled as represented in  FIG. 4 , onto a 96-well plate  70 . 
     The plate  31 , row  41 , column  51 , and diagonal  61  pooled DNA complete with positive and negative controls C 4  and D 4  respectively, are further re-pooling onto the Matrix Pool Plate  FIG. 5 , which for illustrative purposes is a 96-well plate  80 . This combination further narrows the search for the well with the clone of interest.  FIG. 4  depicts the complete set of intermediate subpools in the hierarchical structure plate pool  30 , row pool  40 , column pool  50  and diagonal pool  60  that were generated by processing each individual subpool to extract the necessary material according to the hierarchical structure.  FIG. 4  is where the isolated material from the subpools is stored in a stable form before repooling each intermediate subpooled material into the Final Screening Pools as shown in  FIG. 5 . 
     Material from the complete set of intermediate subpools is further combined and repooled into the Matrix Pool Plate  FIG. 5 , the researcher will then receive two identical Matrix Pool Plates  FIG. 5  for each Superpool to use to perform Round II PCR.  FIG. 5  represents the key step to the present invention.  FIG. 5  represents the repooling of the intermediate subpooled material into a number of final screening pools based on a specific repooling design, wherein individual information is in at least four final screening pools and no more than eight final screening pools. When the plate of final screening pool materials is screened, the specific coordinates are determined which allows the identification of the well position of the clone of interest. 
     EXAMPLE 1 
     Detailed Description of Pools &amp; Superpools: 
     This description is based on 384 well index plates, but it could be used with other plate formats as well with appropriate considerations. It is also based on a BAC genomic DNA library comprised of individual BAC clones, but it could be used with a large variety of biological sample collections or chemical sample collections. The system consists of a collection of multiple Superpools that are screened during First Round PCR, to determine which set of Matrix Pools to screen during Second Round PCR. The total number of Superpools is determined by the total number of clones in the BAC library. Each Superpool has it&#39;s own 96-well plate of corresponding Matrix Pools. 
     Superpools: Each superpool consists of twelve consecutive 384-well plates from a BAC library. DNA is prepared by growing EACH BAC CLONE separately (to avoid growth competition between BAC clones) then combining the 4,608 cultures into one large-scale BAC prep. The Superpool of BAC DNA is then aliquoted onto a 96-well plate. Superpool SP-1 has all the BAC clones in the first twelve plates of the BAC library (Plate 001 to Plate 012). 
     Superpool SP-2 has all the BAC clones in the second twelve plates of the BAC library (Plate 013 to Plate 024). This naming continues for the entire library. 
     Matrix Pools: For each superpool there is one set Matrix Pools (this set of 36 Matrix Pools are aliquoted onto a Matrix Pool Plate. The Matrix Pools of Superpool #1 are named: 
     Matrix Plate Pools 1MPP-A1 through 1MPP-H1 for the 8 wells that contain the matrix of plates 1-12 in Superpool one. Each Matrix Plate Pool contains 1,152 clones. Table 1 indicates the clones in each well. The same process is repeated for as many superpools as are needed for the complete library. 
                                   TABLE 1                   Matrix Plate Pools; Corresponding to FIG. 5, 31.                    clones contained in plate #           Matrix well #   of the specific superpool                   A1   1,2,3           B1   4,5,6           C1   7,8,9           D1   10,11,12           E1   1,5,9           F1   2,6,10           G1   3,7,11           H1   4,8,12                    
Matrix Row Pools 1MRP-A2 through 1MRP-H2 for the 8 wells that contain the matrix of rows A-P in Superpool #1. Each Matrix Row Pool contains 1,152 clones for twelve 384 well plates. See table 2 for the composition of each well in the Matrix Row Pools.
 
                                   TABLE 2                   Matrix Row Pools; Corresponding to FIG. 5, 41.                    Clones contained in row letter           Matrix well #   of the specific superpool                   A2   A,B,C,D           B2   E,F,G,H           C2   I,J,K,L           D2   M,N,O,P           E2   A,E,I,M           F2   B,F,J,N           G2   C,G,K,O           H2   D,H,L,P                    
Matrix Column Pools 1MPP-A3 through 1MPP-B4 for the 10 wells that contain the matrix of columns 1-24 in Superpool #1. See table 3 for the exact composition of each well in the Matrix Column Pools. The Matrix Column Pools in wells A3 through D3 have 1,152 clones (6 different columns X 192 column wells/plate=1,152 clones per Matrix Column Pool). The Matrix Column Pools in wells E3 through B4 contain 768 clones (4 different columns X 192 column wells/plate=768 clones per Matrix Row Pool).
 
                                   TABLE 3                   Matrix Column Pools; Corresponding to FIG. 5, 51.                    Clones contained in column #           Matrix well #   of the specific Superpool                   A3   1,2,3,4,5,6           B3   7,8,9,10,11,12           C3   13,14,15,16,17,18           D3   19,20,21,22,23,24           E3   1,7,13,19           F3   2,8,14,20           G3   3,9,15,21           H3   4,10,16,22           A4   5,11,17,23           B4   6,12,18,24                    
Matrix Diagonal Pools 1MDP-G4 through 1MDP-H5 for the 10 wells that contain the matrix of diagonals 1-24 in Superpool #1. See table 4 for the exact composition of each pool in the Diagonal Pools. The diagonal pools are a collection of clones from all twelve plates in one superpool that has been transected by a plane that goes diagonal in an XY plane and diagonal in a XZ plane through the 12 plates. The diagonals are named by the number of the column that the clone from row A on plate 1 of the specific diagonal. Table 5 shows the exact composition of the Matrix Diagonal Pools. In wells G4 through B5 have 1,152 clones (6 different diagonals X 12 plates/diagonal X 16 column wells/plate=1,152 clones per Matrix Diagonal Pool). The Matrix Diagonal Pools in wells C5 through H5 contain 768 clones (4 different diagonals X 12 plates/diagonal X 16 column wells/plate=768 clones per Matrix Row Pool).
 
     
       
         
               
             
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Diagonal Pool Composition; Depicted graphicallyby FIG. 3D and correspondig 
               
               
                 to the construction of FIG. 4, 60. 
               
             
          
           
               
                 Diagonal 
                 clones contained in the specific superpool labeled by (plate, row, column) (note: as 
               
               
                 pool # 
                 the column gets to 24, it wraps back to column 1 for a 16 row by 24 column plate) 
               
               
                   
               
               
                  1 
                 1A1,1B2,1C3 . . . 1P16; 2A2,2B3,2C4 . . . 2P17; . . . ; 12A12,12B13,12C14 . . . 12P3 
               
               
                  2 
                 1A2,1B3,1C4 . . . 1P17; 2A3,2B4,2C5 . . . 2P18; . . . ; 12A13,12B14,12C15 . . . 12P4 
               
               
                  3 
                 1A3,1B4,1C5 . . . 1P18; 2A4,2B5,2C6 . . . 2P19; . . . ; 12A14,12B15,12C16 . . . 12P5 
               
               
                  4 
                 1A4,1B5,1C6 . . . 1P19; 2A5,2B6,2C7 . . . 2P20; . . . ; 12A15,12B16,12C17 . . . 12P6 
               
               
                  5 
                 1A5,1B6,1C7 . . . 1P20; 2A6,2B7,2C8 . . . 2P21; . . . ; 12A16,12B17,12C18 . . . 12P7 
               
               
                  6 
                 1A6,1B7,1C8 . . . 1P21; 2A7,2B8,2C9 . . . 2P22; . . . ; 12A17,12B18,12C19 . . . 12P8 
               
               
                  7 
                 1A7,1B8,1C9 . . . 1P22; 2A8,2B9,2C10 . . . 2P23; . . . ; 12A18,12B19,12C20 . . . 12P9 
               
               
                  8 
                 1A8,1B9,1C10 . . . 1P23; 2A9,2B10,2C11 . . . 2P24; . . . ; 12A19,12B20,12C21 . . . 12P10 
               
               
                  9 
                 1A9,1B10,1C11 . . . 1P24; 2A10,2B11,2C12 . . . 2P1; . . . ; 12A20,12B21,12C22 . . . 12P11 
               
               
                 10 
                 1A10,1B11,1C12 . . . 1P1; 2A11,2B12,2C13 . . . 2P2; . . . ; 12A21,12B22,12C23 . . . 12P12 
               
               
                 11 
                 1A11,1B12,1C13 . . . 1P2; 2A12,2B13,2C14 . . . 2P3; . . . ; 12A22,12B23,12C24 . . . 12P13 
               
               
                 12 
                 1A12,1B13,1C14 . . . 1P3; 2A13,2B14,2C15 . . . 2P4; . . . ; 12A23,12B24,12C1 . . . 12P14 
               
               
                 13 
                 1A13,1B14,1C15 . . . 1P4; 2A14,2B15,2C16 . . . 2P5; . . . ; 12A24,12B1,12C2 . . . 12P15 
               
               
                 14 
                 1A14,1B15,1C16 . . . 1P5; 2A15,2B16,2C17 . . . 2P6; . . . ; 12A1,12B2,12C3 . . . 12P16 
               
               
                 15 
                 1A15,1B16,1C17 . . . 1P6; 2A16,2B17,2C18 . . . 2P7; . . . ; 12A2,12B3,12C4 . . . 12P17 
               
               
                 16 
                 1A16,1B17,1C18 . . . 1P7; 2A17,2B18,2C19 . . . 2P8; . . . ; 12A3,12B4,12C5 . . . 12P18 
               
               
                 17 
                 1A17,1B18,1C19 . . . 1P8; 2A18,2B19,2C20 . . . 2P9; . . . ; 12A4,12B5,12C6 . . . 12P19 
               
               
                 18 
                 1A18,1B19,1C20 . . . 1P9; 2A19,2B20,2C21 . . . 2P10; . . . ; 12A5,12B6,12C7 . . . 12P20 
               
               
                 19 
                 1A19,1B20,1C21 . . . 1P10; 2A20,2B21,2C22 . . . 2P11; . . . ; 12A6,12B7,12C8 . . . 12P21 
               
               
                 20 
                 1A20,1B21,1C22 . . . 1P11; 2A21,2B22,2C23 . . . 2P12; . . . ; 12A7,12B8,12C9 . . . 12P22 
               
               
                 21 
                 1A21,1B22,1C23 . . . 1P12; 2A22,2B23,2C24 . . . 2P13; . . . ; 12A8,12B9,12C10 . . . 12P23 
               
               
                 22 
                 1A22,1B23,1C24 . . . 1P13; 2A23,2B24,2C1 . . . 2P14; . . . ; 12A9,12B10,12C11 . . . 12P24 
               
               
                 23 
                 1A23,1B24,1C1 . . . 1P14; 2A24,2B1,2C2 . . . 2P15; . . . ; 12A10,12B11,12C12 . . . 12P1 
               
               
                 24 
                 1A24,1B1,1C2 . . . 1P15; 2A1,2B2,2C3 . . . 2P16; . . . ; 12A11,12B12,12C13 . . . 12P2 
               
               
                   
               
             
          
         
       
     
     It is clear that this table is but just an example of a diagonal scheme that is non-redundant with other pools. This table is not limited to one specific diagonal, since there are additional diagonal strategies that can also included as obvious expansions on this diagonal strategy. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Matrix Diagonal Pools; Corresponding to FIG. 5, 61. 
               
             
          
           
               
                   
                   
                 Clones contained in diagonal # 
               
               
                   
                 Matrix well # 
                 of the specific superpool 
               
               
                   
               
               
                   
                 G4 
                 1,2,3,4,5,6 
               
               
                   
                 H4 
                 7,8,9,10,11,12 
               
               
                   
                 A5 
                 13,14,15,16,17,18 
               
               
                   
                 B5 
                 19,20,21,22,23,24 
               
               
                   
                 C5 
                 1,7,13,19 
               
               
                   
                 D5 
                 2,8,14,20 
               
               
                   
                 E5 
                 3,9,15,21 
               
               
                   
                 F5 
                 4,10,16,22 
               
               
                   
                 G5 
                 5,11,17,23 
               
               
                   
                 H5 
                 6,12,18,24 
               
               
                   
               
             
          
         
       
     
     After screening the matrix pools by one of many possible methods, the identity of a specific positive clone from the library can be determined. The specific identification can be determined by a number of ways. If the pool design and matrix design are written or available in electronic form, the unique clone can be identified by a visual or electronic search. There can also be algorithms written based on the pool and matrix designs that can identify the unique clone. 
     The second example describes a method to form a matrix of a variety of different superpools and/or across a variety of different types of pools to allow the screening of the complete library with just one round of experiments. To do this, each small pool or subpool would be added to between 6 and 20 of the collection of re-pooled intermediate or final pools. Then with the total number of pools of between 40 and 180, and more preferably between 80 and 94, the complete library could be screened with high confidence and the ability to resolve multiple hits. If the library had a large redundancy of signal, the total number of pools could be increased to maintain accurate resolving power of the matrix solution. Note: 94 experiments is the preferred number, because current screening technologies are performed on a 96-well index plate format (94 experiments will allow room for a positive control and negative control). 
     In the second example we will teach an additional method that allows the complete library to be screened in one step while still maintaining the resolution of the superpool individual pools formed in Example 1. 
     EXAMPLE 2 
     This example further illustrates and teaches the advantages and possibilities of the current invention. This example is also based on 384 well index plates, but it could be used with other plate formats as well with appropriate considerations. It is also based on a BAC genomic DNA library comprised of individual BAC clones, but it could be used with a large variety of biological collections. The superpools will be composed of eight 384 well plates per superpool and with 10 superpools combined into one large set of matrix pools. Therefore there will be 80 plates (30,720 individual BAC clones in the library) in this one matrix screening that can be tested with a limited number of tests while still maintaining good resolution to an individual clone or may possibly requires screening a few clones during the clone confirmation test directly on the clone(s) of interest. This scheme also allows a single set of experiments (instead of two sets of experiments as described in Example 1). 
     In this scheme, the individual superpools are numbered so that each individual ⅓ plate, row, column and diagonal pool has a unique number. Since there are 88 pools per superpool and ten superpools in this example, there are a total of 880 individual pools that will be combines into one large set of matrix pools. Depending on the number of redundant clones in the BAC library (a function of the genome size and the insert size of the BAC clones), the idealized degree of redundancy can dramatically improve the ability to identify multiple positive clones in one screening and thus minimize ambiguous results (when the user is analyzing data from the screening experiments). 
     The first ⅓ plate pools are formed by collecting all of the clones in plate 1 from columns 1-8. Then the second ⅓ plate pool is all of the clones from columns 9-16 of plate one. This continues on until the 24 th  ⅓ plate pool is from columns 17-24 of plate 8. The twenty-four ⅓ plate pools from superpool two would be considered being in pools 89-112 and so on until the tenth superpool where the ⅓ plate pools would be in pools 793-816. 
     The row pools would be built the same way as Example 1 but since there are only 8 plates in each superpool, each pool would have 192 clones. All of the clones in row A of the eight plates would be pooled together and these clones would be considered pool number 25. This would continue on in a similar fashion so all of the clones in row B of all eight plates of the superpool would belong to pool 26 (and so on) until finally, the pool of all of the clones in row P of the first eight plates would belong to pool number 40. Similarly, the row pools from the second superpool will be in pools numbered 113-128. This would continue in a similar fashion until all of the superpool individual clones belong to row pools and each are assigned unique numbers. 
     The column pools would be formed the same way as in Example 1 but since there are only 8 plates in each superpool, each pool would have 128 clones. All of the clones in column 1 of the eight plates would be pooled together and would belong to pool number 41. This would continue on in a similar fashion until all of the clones in column 2 of all eight plates of the superpool would belong to pool 42 (and so on). Until finally, the pool of all of the clones in column 24 of the first eight plates belong to pool number 64. Similarly, the column pools from the second superpool will be in pools numbered 129-152. This would continue in a similar fashion until all of the superpools belong to column pools and each are assigned unique numbers. 
     The diagonal pools would be formed the same way as in Example 1 but since there are only 8 plates in each superpool, each pool would have 128 clones. See table 6 for the 8 plate superpool diagonal composition. All of the clones in diagonal 1 of the eight plates would be pooled together and would belong to pool number 65. This would continue on in a similar fashion until all of the clones in diagonal 2 of all eight plates of the superpool would belong to pool 66 (and so on). Until finally, the pool of all of the clones in diagonal 24 of the first eight plates belong to pool number 88. Similarly, the diagonal pools from the second superpool will be in pools numbered 152-176. This would continue in a similar fashion until all of the superpools belong to diagonal pools and each are assigned unique numbers. 
     To see one design of many possible schemes for identifying a complete set unique pool numbers, please see Table 7. Table 7 is designed for 88 pools in each subset (superpool) and ten subset (superpools) in the complete set. These unique pool numbers are used to construct various tested screening pool pooling strategies. 
     
       
         
               
             
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Diagonal pool composition for a 8 plate superpool. 
               
             
          
           
               
                 Diagonal 
                 clones contained in the specific superpool labeled by (plate, row, column) (note: as 
               
               
                 pool # 
                 the column gets to 24, it wraps back to column 1 for a 16 row by 24 column plate) 
               
               
                   
               
               
                  1 
                 1A1,1B2,1C3 . . . 1P16; 2A2,2B3,2C4 . . . 2P17; . . . ; 8A8,8B9,8C10 . . . 8P23 
               
               
                  2 
                 1A2,1B3,1C4 . . . 1P17; 2A3,2B4,2C5 . . . 2P18; . . . ; 8A9,8B10,8C11 . . . 8P24 
               
               
                  3 
                 1A3,1B4,1C5 . . . 1P18; 2A4,2B5,2C6 . . . 2P19; . . . ; 8A10,8B11,12C12 . . . 8P1 
               
               
                  4 
                 1A4,1B5,1C6 . . . 1P19; 2A5,2B6,2C7 . . . 2P20; . . . ; 8A11,8B12,8C13 . . . 8P2 
               
               
                  5 
                 1A5,1B6,1C7 . . . 1P20; 2A6,2B7,2C8 . . . 2P21; . . . ; 8A12,8B13,8C14 . . . 8P3 
               
               
                  6 
                 1A6,1B7,1C8 . . . 1P21; 2A7,2B8,2C9 . . . 2P22; . . . ; 8A13,8B14,8C15 . . . 8P4 
               
               
                  7 
                 1A7,1B8,1C9 . . . 1P22; 2A8,2B9,2C10 . . . 2P23; . . . ; 8A14,8B15,8C16 . . . 8P5 
               
               
                  8 
                 1A8,1B9,1C10 . . . 1P23; 2A9,2B10,2C11 . . . 2P24; . . . ; 8A15,8B16,8C17 . . . 8P6 
               
               
                  9 
                 1A9,1B10,1C11 . . . 1P24; 2A10,2B11,2C12 . . . 2P1; . . . ; 8A16,8B17,8C18 . . . 8P7 
               
               
                 10 
                 1A10,1B11,1C12 . . . 1P1; 2A11,2B12,2C13 . . . 2P2; . . . ; 8A17,8B18,8C19 . . . 8P8 
               
               
                 11 
                 1A11,1B12,1C13 . . . 1P2; 2A12,2B13,2C14 . . . 2P3; . . . ; 8A18,8B19,8C20 . . . 8P9 
               
               
                 12 
                 1A12,1B13,1C14 . . . 1P3; 2A13,2B14,2C15 . . . 2P4; . . . ; 8A19,8B20,8C21 . . . 8P10 
               
               
                 13 
                 1A13,1B14,1C15 . . . 1P4; 2A14,2B15,2C16 . . . 2P5; . . . ; 8A20,8B21,8C22 . . . 8P11 
               
               
                 14 
                 1A14,1B15,1C16 . . . 1P5; 2A15,2B16,2C17 . . . 2P6; . . . ; 8A21,8B22,8C23 . . . 8P12 
               
               
                 15 
                 1A15,1B16,1C17 . . . 1P6; 2A16,2B17,2C18 . . . 2P7; . . . ; 8A22,8B23,8C24 . . . 8P13 
               
               
                 16 
                 1A16,1B17,1C18 . . . 1P7; 2A17,2B18,2C19 . . . 2P8; . . . ; 8A23,8B24,8C1 . . . 8P14 
               
               
                 17 
                 1A17,1B18,1C19 . . . 1P8; 2A18,2B19,2C20 . . . 2P9; . . . ; 8A24,8B1,8C2 . . . 8P15 
               
               
                 18 
                 1A18,1B19,1C20 . . . 1P9; 2A19,2B20,2C21 . . . 2P10; . . . ; 8A1,8B2,8C3 . . . 8P16 
               
               
                 19 
                 1A19,1B20,1C21 . . . 1P10; 2A20,2B21,2C22 . . . 2P11; . . . ; 8A2,8B3,8C4 . . . 8P17 
               
               
                 20 
                 1A20,1B21,1C22 . . . 1P11; 2A21,2B22,2C23 . . . 2P12; . . . ; 8A3,8B4,8C5 . . . 8P18 
               
               
                 21 
                 1A21,1B22,1C23 . . . 1P12; 2A22,2B23,2C24 . . . 2P13; . . . ; 8A4,8B5,8C6 . . . 8P19 
               
               
                 22 
                 1A22,1B23,1C24 . . . 1P13; 2A23,2B24,2C1 . . . 2P14; . . . ; 8A5,8B6,8C7 . . . 8P20 
               
               
                 23 
                 1A23,1B24,1C1 . . . 1P14; 2A24,2B1,2C2 . . . 2P15; . . . ; 8A6,8B7,8C8 . . . 8P21 
               
               
                 24 
                 1A24,1B1,1C2 . . . 1P15; 2A1,2B2,2C3 . . . 2P16; . . . ; 8A7,8B8,8C9 . . . 8P22 
               
               
                   
               
             
          
         
       
     
     Table 7 sequentially assigns numbers to individual small pools or subpools from ten consecutive from eight plates so that the subpools may be repooled into final screening pools according to example alternative embodiments depicted in tables 8-11. 
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Unique pool numbers for the 1/3 plate, row, column and diagonal pools 
               
               
                 of the first ten superpools. 
               
             
          
           
               
                 Individual 
                 Unique pool numbers for 8 plate superpools 1 
               
               
                 superpool 
                 through 10. 
               
             
          
           
               
                 contents 
                 1 
                 2 
                 3 
                 4 
                 5 
                 6 
                 7 
                 8 
                 9 
                 10 
               
               
                   
               
             
          
           
               
                 1/3 plate 1 
                 1 
                 89 
                 177 
                 265 
                 353 
                 441 
                 529 
                 617 
                 705 
                 793 
               
               
                 1/3 plate 2 
                 2 
                 90 
                 178 
                 266 
                 354 
                 442 
                 530 
                 618 
                 706 
                 794 
               
               
                 1/3 plate 3 
                 3 
                 91 
                 179 
                 267 
                 355 
                 443 
                 531 
                 619 
                 707 
                 795 
               
               
                 1/3 plate 4 
                 4 
                 92 
                 180 
                 268 
                 356 
                 444 
                 532 
                 620 
                 708 
                 796 
               
               
                 1/3 plate 5 
                 5 
                 93 
                 181 
                 269 
                 357 
                 445 
                 533 
                 621 
                 709 
                 797 
               
               
                 1/3 plate 6 
                 6 
                 94 
                 182 
                 270 
                 358 
                 446 
                 534 
                 622 
                 710 
                 798 
               
               
                 1/3 plate 7 
                 7 
                 95 
                 183 
                 271 
                 359 
                 447 
                 535 
                 623 
                 711 
                 799 
               
               
                 1/3 plate 8 
                 8 
                 96 
                 184 
                 272 
                 360 
                 448 
                 536 
                 624 
                 712 
                 800 
               
               
                 1/3 plate 9 
                 9 
                 97 
                 185 
                 273 
                 361 
                 449 
                 537 
                 625 
                 713 
                 801 
               
               
                 1/3 plate 10 
                 10 
                 98 
                 186 
                 274 
                 362 
                 450 
                 538 
                 626 
                 714 
                 802 
               
               
                 1/3 plate 11 
                 11 
                 99 
                 187 
                 275 
                 363 
                 451 
                 539 
                 627 
                 715 
                 803 
               
               
                 1/3 plate 12 
                 12 
                 100 
                 188 
                 276 
                 364 
                 452 
                 540 
                 628 
                 716 
                 804 
               
               
                 1/3 plate 13 
                 13 
                 101 
                 189 
                 277 
                 365 
                 453 
                 541 
                 629 
                 717 
                 805 
               
               
                 1/3 plate 14 
                 14 
                 102 
                 190 
                 278 
                 366 
                 454 
                 542 
                 630 
                 718 
                 806 
               
               
                 1/3 plate 15 
                 15 
                 103 
                 191 
                 279 
                 367 
                 455 
                 543 
                 631 
                 719 
                 807 
               
               
                 1/3 plate 16 
                 16 
                 104 
                 192 
                 280 
                 368 
                 456 
                 544 
                 632 
                 720 
                 808 
               
               
                 1/3 plate 17 
                 17 
                 105 
                 193 
                 281 
                 369 
                 457 
                 545 
                 633 
                 721 
                 809 
               
               
                 1/3 plate 18 
                 18 
                 106 
                 194 
                 282 
                 370 
                 458 
                 546 
                 634 
                 722 
                 810 
               
               
                 1/3 plate 19 
                 19 
                 107 
                 195 
                 283 
                 371 
                 459 
                 547 
                 635 
                 723 
                 811 
               
               
                 1/3 plate 20 
                 20 
                 108 
                 196 
                 284 
                 372 
                 460 
                 548 
                 636 
                 724 
                 812 
               
               
                 1/3 plate 21 
                 21 
                 109 
                 197 
                 285 
                 373 
                 461 
                 549 
                 637 
                 725 
                 813 
               
               
                 1/3 plate 22 
                 22 
                 110 
                 198 
                 286 
                 374 
                 462 
                 550 
                 638 
                 726 
                 814 
               
               
                 1/3 plate 23 
                 23 
                 111 
                 199 
                 287 
                 375 
                 463 
                 551 
                 639 
                 727 
                 815 
               
               
                 1/3 plate 24 
                 24 
                 112 
                 200 
                 288 
                 376 
                 464 
                 552 
                 640 
                 728 
                 816 
               
               
                 row A 
                 25 
                 113 
                 201 
                 289 
                 377 
                 465 
                 553 
                 641 
                 729 
                 817 
               
               
                 row B 
                 26 
                 114 
                 202 
                 290 
                 378 
                 466 
                 554 
                 642 
                 730 
                 818 
               
               
                 row C 
                 27 
                 115 
                 203 
                 291 
                 379 
                 467 
                 555 
                 643 
                 731 
                 819 
               
               
                 row D 
                 28 
                 116 
                 204 
                 292 
                 380 
                 468 
                 556 
                 644 
                 732 
                 820 
               
               
                 row E 
                 29 
                 117 
                 205 
                 293 
                 381 
                 469 
                 557 
                 645 
                 733 
                 821 
               
               
                 row F 
                 30 
                 118 
                 206 
                 294 
                 382 
                 470 
                 558 
                 646 
                 734 
                 822 
               
               
                 row G 
                 31 
                 119 
                 207 
                 295 
                 383 
                 471 
                 559 
                 647 
                 735 
                 823 
               
               
                 row H 
                 32 
                 120 
                 208 
                 296 
                 384 
                 472 
                 560 
                 648 
                 736 
                 824 
               
               
                 row I 
                 33 
                 121 
                 209 
                 297 
                 385 
                 473 
                 561 
                 649 
                 737 
                 825 
               
               
                 row J 
                 34 
                 122 
                 210 
                 298 
                 386 
                 474 
                 562 
                 650 
                 738 
                 826 
               
               
                 row K 
                 35 
                 123 
                 211 
                 299 
                 387 
                 475 
                 563 
                 651 
                 739 
                 827 
               
               
                 row L 
                 36 
                 124 
                 212 
                 300 
                 388 
                 476 
                 564 
                 652 
                 740 
                 828 
               
               
                 row M 
                 37 
                 125 
                 213 
                 301 
                 389 
                 477 
                 565 
                 653 
                 741 
                 829 
               
               
                 row N 
                 38 
                 126 
                 214 
                 302 
                 390 
                 478 
                 566 
                 654 
                 742 
                 830 
               
               
                 row O 
                 39 
                 127 
                 215 
                 303 
                 391 
                 479 
                 567 
                 655 
                 743 
                 831 
               
               
                 row P 
                 40 
                 128 
                 216 
                 304 
                 392 
                 480 
                 568 
                 656 
                 744 
                 832 
               
               
                 column 1 
                 41 
                 129 
                 217 
                 305 
                 393 
                 481 
                 569 
                 657 
                 745 
                 833 
               
               
                 column 2 
                 42 
                 130 
                 218 
                 306 
                 394 
                 482 
                 570 
                 658 
                 746 
                 834 
               
               
                 column 3 
                 43 
                 131 
                 219 
                 307 
                 395 
                 483 
                 571 
                 659 
                 747 
                 835 
               
               
                 column 4 
                 44 
                 132 
                 220 
                 308 
                 396 
                 484 
                 572 
                 660 
                 748 
                 836 
               
               
                 column 5 
                 45 
                 133 
                 221 
                 309 
                 397 
                 485 
                 573 
                 661 
                 749 
                 837 
               
               
                 column 6 
                 46 
                 134 
                 222 
                 310 
                 398 
                 486 
                 574 
                 662 
                 750 
                 838 
               
               
                 column 7 
                 47 
                 135 
                 223 
                 311 
                 399 
                 487 
                 575 
                 663 
                 751 
                 839 
               
               
                 column 8 
                 48 
                 136 
                 224 
                 312 
                 400 
                 488 
                 576 
                 664 
                 752 
                 840 
               
               
                 column 9 
                 49 
                 137 
                 225 
                 313 
                 401 
                 489 
                 577 
                 665 
                 753 
                 841 
               
               
                 column 10 
                 50 
                 138 
                 226 
                 314 
                 402 
                 490 
                 578 
                 666 
                 754 
                 842 
               
               
                 column 11 
                 51 
                 139 
                 227 
                 315 
                 403 
                 491 
                 579 
                 667 
                 755 
                 843 
               
               
                 column 12 
                 52 
                 140 
                 228 
                 316 
                 404 
                 492 
                 580 
                 668 
                 756 
                 844 
               
               
                 column 13 
                 53 
                 141 
                 229 
                 317 
                 405 
                 493 
                 581 
                 669 
                 757 
                 845 
               
               
                 column 14 
                 54 
                 142 
                 230 
                 318 
                 406 
                 494 
                 582 
                 670 
                 758 
                 846 
               
               
                 column 15 
                 55 
                 143 
                 231 
                 319 
                 407 
                 495 
                 583 
                 671 
                 759 
                 847 
               
               
                 column 16 
                 56 
                 144 
                 232 
                 320 
                 408 
                 496 
                 584 
                 672 
                 760 
                 848 
               
               
                 column 17 
                 57 
                 145 
                 233 
                 321 
                 409 
                 497 
                 585 
                 673 
                 761 
                 849 
               
               
                 column 18 
                 58 
                 146 
                 234 
                 322 
                 410 
                 498 
                 586 
                 674 
                 762 
                 850 
               
               
                 column 19 
                 59 
                 147 
                 235 
                 323 
                 411 
                 499 
                 587 
                 675 
                 763 
                 851 
               
               
                 column 20 
                 60 
                 148 
                 236 
                 324 
                 412 
                 500 
                 588 
                 676 
                 764 
                 852 
               
               
                 column 21 
                 61 
                 149 
                 237 
                 325 
                 413 
                 501 
                 589 
                 677 
                 765 
                 853 
               
               
                 column 22 
                 62 
                 150 
                 238 
                 326 
                 414 
                 502 
                 590 
                 678 
                 766 
                 854 
               
               
                 column 23 
                 63 
                 151 
                 239 
                 327 
                 415 
                 503 
                 591 
                 679 
                 767 
                 855 
               
               
                 column 24 
                 64 
                 152 
                 240 
                 328 
                 416 
                 504 
                 592 
                 680 
                 768 
                 856 
               
               
                 diagonal 1 
                 65 
                 153 
                 241 
                 329 
                 417 
                 505 
                 593 
                 681 
                 769 
                 857 
               
               
                 diagonal 2 
                 66 
                 154 
                 242 
                 330 
                 418 
                 506 
                 594 
                 682 
                 770 
                 858 
               
               
                 diagonal 3 
                 67 
                 155 
                 243 
                 331 
                 419 
                 507 
                 595 
                 683 
                 771 
                 859 
               
               
                 diagonal 4 
                 68 
                 156 
                 244 
                 332 
                 420 
                 508 
                 596 
                 684 
                 772 
                 860 
               
               
                 diagonal 5 
                 69 
                 157 
                 245 
                 333 
                 421 
                 509 
                 597 
                 685 
                 773 
                 861 
               
               
                 diagonal 6 
                 70 
                 158 
                 246 
                 334 
                 422 
                 510 
                 598 
                 686 
                 774 
                 862 
               
               
                 diagonal 7 
                 71 
                 159 
                 247 
                 335 
                 423 
                 511 
                 599 
                 687 
                 775 
                 863 
               
               
                 diagonal 8 
                 72 
                 160 
                 248 
                 336 
                 424 
                 512 
                 600 
                 688 
                 776 
                 864 
               
               
                 diagonal 9 
                 73 
                 161 
                 249 
                 337 
                 425 
                 513 
                 601 
                 689 
                 777 
                 865 
               
               
                 diagonal 10 
                 74 
                 162 
                 250 
                 338 
                 426 
                 514 
                 602 
                 690 
                 778 
                 866 
               
               
                 diagonal 11 
                 75 
                 163 
                 251 
                 339 
                 427 
                 515 
                 603 
                 691 
                 779 
                 867 
               
               
                 diagonal 12 
                 76 
                 164 
                 252 
                 340 
                 428 
                 516 
                 604 
                 692 
                 780 
                 868 
               
               
                 diagonal 13 
                 77 
                 165 
                 253 
                 341 
                 429 
                 517 
                 605 
                 693 
                 781 
                 869 
               
               
                 diagonal 14 
                 78 
                 166 
                 254 
                 342 
                 430 
                 518 
                 606 
                 694 
                 782 
                 870 
               
               
                 diagonal 15 
                 79 
                 167 
                 255 
                 343 
                 431 
                 519 
                 607 
                 695 
                 783 
                 871 
               
               
                 diagonal 16 
                 80 
                 168 
                 256 
                 344 
                 432 
                 520 
                 608 
                 696 
                 784 
                 872 
               
               
                 diagonal 17 
                 81 
                 169 
                 257 
                 345 
                 433 
                 521 
                 609 
                 697 
                 785 
                 873 
               
               
                 diagonal 18 
                 82 
                 170 
                 258 
                 346 
                 434 
                 522 
                 610 
                 698 
                 786 
                 874 
               
               
                 diagonal 19 
                 83 
                 171 
                 259 
                 347 
                 435 
                 523 
                 611 
                 699 
                 787 
                 875 
               
               
                 diagonal 20 
                 84 
                 172 
                 260 
                 348 
                 436 
                 524 
                 612 
                 700 
                 788 
                 876 
               
               
                 diagonal 21 
                 85 
                 173 
                 261 
                 349 
                 437 
                 525 
                 613 
                 701 
                 789 
                 877 
               
               
                 diagonal 22 
                 86 
                 174 
                 262 
                 350 
                 438 
                 526 
                 614 
                 702 
                 790 
                 878 
               
               
                 diagonal 23 
                 87 
                 175 
                 263 
                 351 
                 439 
                 527 
                 615 
                 703 
                 791 
                 879 
               
               
                 diagonal 24 
                 88 
                 176 
                 264 
                 352 
                 440 
                 528 
                 616 
                 704 
                 792 
                 880 
               
               
                   
               
             
          
         
       
     
     Tables 8-11 describe various embodiments in the systematic or randomization of the loading of the small pool or subpool plate, row, column, and diagonal pooled DNA ( FIG. 4 ) into an alternate Matrix Pool Plate format ( FIG.5 ). 
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 Example 3 screening pool design. 
               
               
                 94 seq 5 screening pool 
               
               
                 design 
               
             
          
           
               
                 Screening 
                   
               
               
                 pool # 
                 Unique pools contained in each screening pool 
               
               
                   
               
             
          
           
               
                 1 
                 1 
                 95 
                 189 
                 283 
                 377 
                 471 
                 565 
                 659 
                 753 
                 847 
                 301 
                 827 
               
               
                 2 
                 2 
                 96 
                 190 
                 284 
                 378 
                 472 
                 566 
                 660 
                 754 
                 848 
                 302 
                 828 
               
               
                 3 
                 3 
                 97 
                 191 
                 285 
                 379 
                 473 
                 567 
                 661 
                 755 
                 849 
                 303 
                 829 
               
               
                 4 
                 4 
                 98 
                 192 
                 286 
                 380 
                 474 
                 568 
                 662 
                 756 
                 850 
                 304 
                 830 
               
               
                 5 
                 5 
                 99 
                 193 
                 287 
                 381 
                 475 
                 569 
                 663 
                 757 
                 851 
                 377 
                 831 
               
               
                 6 
                 6 
                 100 
                 194 
                 288 
                 382 
                 476 
                 570 
                 664 
                 758 
                 852 
                 378 
                 832 
               
               
                 7 
                 7 
                 101 
                 195 
                 289 
                 383 
                 477 
                 571 
                 665 
                 759 
                 853 
                 379 
               
               
                 8 
                 8 
                 102 
                 196 
                 290 
                 384 
                 478 
                 572 
                 666 
                 760 
                 854 
                 380 
               
               
                 9 
                 9 
                 103 
                 197 
                 291 
                 385 
                 479 
                 573 
                 667 
                 761 
                 855 
                 381 
               
               
                 10 
                 10 
                 104 
                 198 
                 292 
                 386 
                 480 
                 574 
                 668 
                 762 
                 856 
                 382 
               
               
                 11 
                 11 
                 105 
                 199 
                 293 
                 387 
                 481 
                 575 
                 669 
                 763 
                 857 
                 383 
               
               
                 12 
                 12 
                 106 
                 200 
                 294 
                 388 
                 482 
                 576 
                 670 
                 764 
                 858 
                 384 
               
               
                 13 
                 13 
                 107 
                 201 
                 295 
                 389 
                 483 
                 577 
                 671 
                 765 
                 859 
                 385 
               
               
                 14 
                 14 
                 108 
                 202 
                 296 
                 390 
                 484 
                 578 
                 672 
                 766 
                 860 
                 386 
               
               
                 15 
                 15 
                 109 
                 203 
                 297 
                 391 
                 485 
                 579 
                 673 
                 767 
                 861 
                 387 
               
               
                 16 
                 16 
                 110 
                 204 
                 298 
                 392 
                 486 
                 580 
                 674 
                 768 
                 862 
                 388 
               
               
                 17 
                 17 
                 111 
                 205 
                 299 
                 393 
                 487 
                 581 
                 675 
                 769 
                 863 
                 389 
               
               
                 18 
                 18 
                 112 
                 206 
                 300 
                 394 
                 488 
                 582 
                 676 
                 770 
                 864 
                 390 
               
               
                 19 
                 19 
                 113 
                 207 
                 301 
                 395 
                 489 
                 583 
                 677 
                 771 
                 865 
                 391 
               
               
                 20 
                 20 
                 114 
                 208 
                 302 
                 396 
                 490 
                 584 
                 678 
                 772 
                 866 
                 392 
               
               
                 21 
                 21 
                 115 
                 209 
                 303 
                 397 
                 491 
                 585 
                 679 
                 773 
                 867 
                 465 
               
               
                 22 
                 22 
                 116 
                 210 
                 304 
                 398 
                 492 
                 586 
                 680 
                 774 
                 868 
                 466 
               
               
                 23 
                 23 
                 117 
                 211 
                 305 
                 399 
                 493 
                 587 
                 681 
                 775 
                 869 
                 467 
               
               
                 24 
                 24 
                 118 
                 212 
                 306 
                 400 
                 494 
                 588 
                 682 
                 776 
                 870 
                 468 
               
               
                 25 
                 25 
                 119 
                 213 
                 307 
                 401 
                 495 
                 589 
                 683 
                 777 
                 871 
                 469 
               
               
                 26 
                 26 
                 120 
                 214 
                 308 
                 402 
                 496 
                 590 
                 684 
                 778 
                 872 
                 470 
               
               
                 27 
                 27 
                 121 
                 215 
                 309 
                 403 
                 497 
                 591 
                 685 
                 779 
                 873 
                 471 
               
               
                 28 
                 28 
                 122 
                 216 
                 310 
                 404 
                 498 
                 592 
                 686 
                 780 
                 874 
                 472 
               
               
                 29 
                 29 
                 123 
                 217 
                 311 
                 405 
                 499 
                 593 
                 687 
                 781 
                 875 
                 473 
               
               
                 30 
                 30 
                 124 
                 218 
                 312 
                 406 
                 500 
                 594 
                 688 
                 782 
                 876 
                 474 
               
               
                 31 
                 31 
                 125 
                 219 
                 313 
                 407 
                 501 
                 595 
                 689 
                 783 
                 877 
                 475 
               
               
                 32 
                 32 
                 126 
                 220 
                 314 
                 408 
                 502 
                 596 
                 690 
                 784 
                 878 
                 476 
               
               
                 33 
                 33 
                 127 
                 221 
                 315 
                 409 
                 503 
                 597 
                 691 
                 785 
                 879 
                 477 
               
               
                 34 
                 34 
                 128 
                 222 
                 316 
                 410 
                 504 
                 598 
                 692 
                 786 
                 880 
                 478 
               
               
                 35 
                 35 
                 129 
                 223 
                 317 
                 411 
                 505 
                 599 
                 693 
                 787 
                 25 
                 479 
               
               
                 36 
                 36 
                 130 
                 224 
                 318 
                 412 
                 506 
                 600 
                 694 
                 788 
                 26 
                 480 
               
               
                 37 
                 37 
                 131 
                 225 
                 319 
                 413 
                 507 
                 601 
                 695 
                 789 
                 27 
                 553 
               
               
                 38 
                 38 
                 132 
                 226 
                 320 
                 414 
                 508 
                 602 
                 696 
                 790 
                 28 
                 554 
               
               
                 39 
                 39 
                 133 
                 227 
                 321 
                 415 
                 509 
                 603 
                 697 
                 791 
                 29 
                 555 
               
               
                 40 
                 40 
                 134 
                 228 
                 322 
                 416 
                 510 
                 604 
                 698 
                 792 
                 30 
                 556 
               
               
                 41 
                 41 
                 135 
                 229 
                 323 
                 417 
                 511 
                 605 
                 699 
                 793 
                 31 
                 557 
               
               
                 42 
                 42 
                 136 
                 230 
                 324 
                 418 
                 512 
                 606 
                 700 
                 794 
                 32 
                 558 
               
               
                 43 
                 43 
                 137 
                 231 
                 325 
                 419 
                 513 
                 607 
                 701 
                 795 
                 33 
                 559 
               
               
                 44 
                 44 
                 138 
                 232 
                 326 
                 420 
                 514 
                 608 
                 702 
                 796 
                 34 
                 560 
               
               
                 45 
                 45 
                 139 
                 233 
                 327 
                 421 
                 515 
                 609 
                 703 
                 797 
                 35 
                 561 
               
               
                 46 
                 46 
                 140 
                 234 
                 328 
                 422 
                 516 
                 610 
                 704 
                 798 
                 36 
                 562 
               
               
                 47 
                 47 
                 141 
                 235 
                 329 
                 423 
                 517 
                 611 
                 705 
                 799 
                 37 
                 563 
               
               
                 48 
                 48 
                 142 
                 236 
                 330 
                 424 
                 518 
                 612 
                 706 
                 800 
                 38 
                 564 
               
               
                 49 
                 49 
                 143 
                 237 
                 331 
                 425 
                 519 
                 613 
                 707 
                 801 
                 39 
                 565 
               
               
                 50 
                 50 
                 144 
                 238 
                 332 
                 426 
                 520 
                 614 
                 708 
                 802 
                 40 
                 566 
               
               
                 51 
                 51 
                 145 
                 239 
                 333 
                 427 
                 521 
                 615 
                 709 
                 803 
                 113 
                 567 
               
               
                 52 
                 52 
                 146 
                 240 
                 334 
                 428 
                 522 
                 616 
                 710 
                 804 
                 114 
                 568 
               
               
                 53 
                 53 
                 147 
                 241 
                 335 
                 429 
                 523 
                 617 
                 711 
                 805 
                 115 
                 641 
               
               
                 54 
                 54 
                 148 
                 242 
                 336 
                 430 
                 524 
                 618 
                 712 
                 806 
                 116 
                 642 
               
               
                 55 
                 55 
                 149 
                 243 
                 337 
                 431 
                 525 
                 619 
                 713 
                 807 
                 117 
                 643 
               
               
                 56 
                 56 
                 150 
                 244 
                 338 
                 432 
                 526 
                 620 
                 714 
                 808 
                 118 
                 644 
               
               
                 57 
                 57 
                 151 
                 245 
                 339 
                 433 
                 527 
                 621 
                 715 
                 809 
                 119 
                 645 
               
               
                 58 
                 58 
                 152 
                 246 
                 340 
                 434 
                 528 
                 622 
                 716 
                 810 
                 120 
                 646 
               
               
                 59 
                 59 
                 153 
                 247 
                 341 
                 435 
                 529 
                 623 
                 717 
                 811 
                 121 
                 647 
               
               
                 60 
                 60 
                 154 
                 248 
                 342 
                 436 
                 530 
                 624 
                 718 
                 812 
                 122 
                 648 
               
               
                 61 
                 61 
                 155 
                 249 
                 343 
                 437 
                 531 
                 625 
                 719 
                 813 
                 123 
                 649 
               
               
                 62 
                 62 
                 156 
                 250 
                 344 
                 438 
                 532 
                 626 
                 720 
                 814 
                 124 
                 650 
               
               
                 63 
                 63 
                 157 
                 251 
                 345 
                 439 
                 533 
                 627 
                 721 
                 815 
                 125 
                 651 
               
               
                 64 
                 64 
                 158 
                 252 
                 346 
                 440 
                 534 
                 628 
                 722 
                 816 
                 126 
                 652 
               
               
                 65 
                 65 
                 159 
                 253 
                 347 
                 441 
                 535 
                 629 
                 723 
                 817 
                 127 
                 653 
               
               
                 66 
                 66 
                 160 
                 254 
                 348 
                 442 
                 536 
                 630 
                 724 
                 818 
                 128 
                 654 
               
               
                 67 
                 67 
                 161 
                 255 
                 349 
                 443 
                 537 
                 631 
                 725 
                 819 
                 201 
                 655 
               
               
                 68 
                 68 
                 162 
                 256 
                 350 
                 444 
                 538 
                 632 
                 726 
                 820 
                 202 
                 656 
               
               
                 69 
                 69 
                 163 
                 257 
                 351 
                 445 
                 539 
                 633 
                 727 
                 821 
                 203 
                 729 
               
               
                 70 
                 70 
                 164 
                 258 
                 352 
                 446 
                 540 
                 634 
                 728 
                 822 
                 204 
                 730 
               
               
                 71 
                 71 
                 165 
                 259 
                 353 
                 447 
                 541 
                 635 
                 729 
                 823 
                 205 
                 731 
               
               
                 72 
                 72 
                 166 
                 260 
                 354 
                 448 
                 542 
                 636 
                 730 
                 824 
                 206 
                 732 
               
               
                 73 
                 73 
                 167 
                 261 
                 355 
                 449 
                 543 
                 637 
                 731 
                 825 
                 207 
                 733 
               
               
                 74 
                 74 
                 168 
                 262 
                 356 
                 450 
                 544 
                 638 
                 732 
                 826 
                 208 
                 734 
               
               
                 75 
                 75 
                 169 
                 263 
                 357 
                 451 
                 545 
                 639 
                 733 
                 827 
                 209 
                 735 
               
               
                 76 
                 76 
                 170 
                 264 
                 358 
                 452 
                 546 
                 640 
                 734 
                 828 
                 210 
                 736 
               
               
                 77 
                 77 
                 171 
                 265 
                 359 
                 453 
                 547 
                 641 
                 735 
                 829 
                 211 
                 737 
               
               
                 78 
                 78 
                 172 
                 266 
                 360 
                 454 
                 548 
                 642 
                 736 
                 830 
                 212 
                 738 
               
               
                 79 
                 79 
                 173 
                 267 
                 361 
                 455 
                 549 
                 643 
                 737 
                 831 
                 213 
                 739 
               
               
                 80 
                 80 
                 174 
                 268 
                 362 
                 456 
                 550 
                 644 
                 738 
                 832 
                 214 
                 740 
               
               
                 81 
                 81 
                 175 
                 269 
                 363 
                 457 
                 551 
                 645 
                 739 
                 833 
                 215 
                 741 
               
               
                 82 
                 82 
                 176 
                 270 
                 364 
                 458 
                 552 
                 646 
                 740 
                 834 
                 216 
                 742 
               
               
                 83 
                 83 
                 177 
                 271 
                 365 
                 459 
                 553 
                 647 
                 741 
                 835 
                 289 
                 743 
               
               
                 84 
                 84 
                 178 
                 272 
                 366 
                 460 
                 554 
                 648 
                 742 
                 836 
                 290 
                 744 
               
               
                 85 
                 85 
                 179 
                 273 
                 367 
                 461 
                 555 
                 649 
                 743 
                 837 
                 291 
                 817 
               
               
                 86 
                 86 
                 180 
                 274 
                 368 
                 462 
                 556 
                 650 
                 744 
                 838 
                 292 
                 818 
               
               
                 87 
                 87 
                 181 
                 275 
                 369 
                 463 
                 557 
                 651 
                 745 
                 839 
                 293 
                 819 
               
               
                 88 
                 88 
                 182 
                 276 
                 370 
                 464 
                 558 
                 652 
                 746 
                 840 
                 294 
                 820 
               
               
                 89 
                 89 
                 183 
                 277 
                 371 
                 465 
                 559 
                 653 
                 747 
                 841 
                 295 
                 821 
               
               
                 90 
                 90 
                 184 
                 278 
                 372 
                 466 
                 560 
                 654 
                 748 
                 842 
                 296 
                 822 
               
               
                 91 
                 91 
                 185 
                 279 
                 373 
                 467 
                 561 
                 655 
                 749 
                 843 
                 297 
                 823 
               
               
                 92 
                 92 
                 186 
                 280 
                 374 
                 468 
                 562 
                 656 
                 750 
                 844 
                 298 
                 824 
               
               
                 93 
                 93 
                 187 
                 281 
                 375 
                 469 
                 563 
                 657 
                 751 
                 845 
                 299 
                 825 
               
               
                 94 
                 94 
                 188 
                 282 
                 376 
                 470 
                 564 
                 658 
                 752 
                 846 
                 300 
                 826 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 Example 4 screening pool design. 
               
               
                 94 seq 4 &amp; SP screening pool 
               
               
                 design 
               
             
          
           
               
                 Screening 
                   
               
               
                 pool # 
                 Unique pools contained in each screening pool 
               
               
                   
               
             
          
           
               
                 1 
                 1 
                 85 
                 169 
                 253 
                 337 
                 421 
                 505 
                 589 
                 673 
                 757 
                 841 
                   
                   
                   
                   
                   
               
               
                 2 
                 2 
                 86 
                 170 
                 254 
                 338 
                 422 
                 506 
                 590 
                 674 
                 758 
                 842 
               
               
                 3 
                 3 
                 87 
                 171 
                 255 
                 339 
                 423 
                 507 
                 591 
                 675 
                 759 
                 843 
               
               
                 4 
                 4 
                 88 
                 172 
                 256 
                 340 
                 424 
                 508 
                 592 
                 676 
                 760 
                 844 
               
               
                 5 
                 5 
                 89 
                 173 
                 257 
                 341 
                 425 
                 509 
                 593 
                 677 
                 761 
                 845 
               
               
                 6 
                 6 
                 90 
                 174 
                 258 
                 342 
                 426 
                 510 
                 594 
                 678 
                 762 
                 846 
               
               
                 7 
                 7 
                 91 
                 175 
                 259 
                 343 
                 427 
                 511 
                 595 
                 679 
                 763 
                 847 
               
               
                 8 
                 8 
                 92 
                 176 
                 260 
                 344 
                 428 
                 512 
                 596 
                 680 
                 764 
                 848 
               
               
                 9 
                 9 
                 93 
                 177 
                 261 
                 345 
                 429 
                 513 
                 597 
                 681 
                 765 
                 849 
               
               
                 10 
                 10 
                 94 
                 178 
                 262 
                 346 
                 430 
                 514 
                 598 
                 682 
                 766 
                 850 
               
               
                 11 
                 11 
                 95 
                 179 
                 263 
                 347 
                 431 
                 515 
                 599 
                 683 
                 767 
                 851 
               
               
                 12 
                 12 
                 96 
                 180 
                 264 
                 348 
                 432 
                 516 
                 600 
                 684 
                 768 
                 852 
               
               
                 13 
                 13 
                 97 
                 181 
                 265 
                 349 
                 433 
                 517 
                 601 
                 685 
                 769 
                 853 
               
               
                 14 
                 14 
                 98 
                 182 
                 266 
                 350 
                 434 
                 518 
                 602 
                 686 
                 770 
                 854 
               
               
                 15 
                 15 
                 99 
                 183 
                 267 
                 351 
                 435 
                 519 
                 603 
                 687 
                 771 
                 855 
               
               
                 16 
                 16 
                 100 
                 184 
                 268 
                 352 
                 436 
                 520 
                 604 
                 688 
                 772 
                 856 
               
               
                 17 
                 17 
                 101 
                 185 
                 269 
                 353 
                 437 
                 521 
                 605 
                 689 
                 773 
                 857 
               
               
                 18 
                 18 
                 102 
                 186 
                 270 
                 354 
                 438 
                 522 
                 606 
                 690 
                 774 
                 858 
               
               
                 19 
                 19 
                 103 
                 187 
                 271 
                 355 
                 439 
                 523 
                 607 
                 691 
                 775 
                 859 
               
               
                 20 
                 20 
                 104 
                 188 
                 272 
                 356 
                 440 
                 524 
                 608 
                 692 
                 776 
                 860 
               
               
                 21 
                 21 
                 105 
                 189 
                 273 
                 357 
                 441 
                 525 
                 609 
                 693 
                 777 
                 861 
               
               
                 22 
                 22 
                 106 
                 190 
                 274 
                 358 
                 442 
                 526 
                 610 
                 694 
                 778 
                 862 
               
               
                 23 
                 23 
                 107 
                 191 
                 275 
                 359 
                 443 
                 527 
                 611 
                 695 
                 779 
                 863 
               
               
                 24 
                 24 
                 108 
                 192 
                 276 
                 360 
                 444 
                 528 
                 612 
                 696 
                 780 
                 864 
               
               
                 25 
                 25 
                 109 
                 193 
                 277 
                 361 
                 445 
                 529 
                 613 
                 697 
                 781 
                 865 
               
               
                 26 
                 26 
                 110 
                 194 
                 278 
                 362 
                 446 
                 530 
                 614 
                 698 
                 782 
                 866 
               
               
                 27 
                 27 
                 111 
                 195 
                 279 
                 363 
                 447 
                 531 
                 615 
                 699 
                 783 
                 867 
               
               
                 28 
                 28 
                 112 
                 196 
                 280 
                 364 
                 448 
                 532 
                 616 
                 700 
                 784 
                 868 
               
               
                 29 
                 29 
                 113 
                 197 
                 281 
                 365 
                 449 
                 533 
                 617 
                 701 
                 785 
                 869 
               
               
                 30 
                 30 
                 114 
                 198 
                 282 
                 366 
                 450 
                 534 
                 618 
                 702 
                 786 
                 870 
               
               
                 31 
                 31 
                 115 
                 199 
                 283 
                 367 
                 451 
                 535 
                 619 
                 703 
                 787 
                 871 
               
               
                 32 
                 32 
                 116 
                 200 
                 284 
                 368 
                 452 
                 536 
                 620 
                 704 
                 788 
                 872 
               
               
                 33 
                 33 
                 117 
                 201 
                 285 
                 369 
                 453 
                 537 
                 621 
                 705 
                 789 
                 873 
               
               
                 34 
                 34 
                 118 
                 202 
                 286 
                 370 
                 454 
                 538 
                 622 
                 706 
                 790 
                 874 
               
               
                 35 
                 35 
                 119 
                 203 
                 287 
                 371 
                 455 
                 539 
                 623 
                 707 
                 791 
                 875 
               
               
                 36 
                 36 
                 120 
                 204 
                 288 
                 372 
                 456 
                 540 
                 624 
                 708 
                 792 
                 876 
               
               
                 37 
                 37 
                 121 
                 205 
                 289 
                 373 
                 457 
                 541 
                 625 
                 709 
                 793 
                 877 
               
               
                 38 
                 38 
                 122 
                 206 
                 290 
                 374 
                 458 
                 542 
                 626 
                 710 
                 794 
                 878 
               
               
                 39 
                 39 
                 123 
                 207 
                 291 
                 375 
                 459 
                 543 
                 627 
                 711 
                 795 
                 879 
               
               
                 40 
                 40 
                 124 
                 208 
                 292 
                 376 
                 460 
                 544 
                 628 
                 712 
                 796 
                 880 
               
               
                 41 
                 41 
                 125 
                 209 
                 293 
                 377 
                 461 
                 545 
                 629 
                 713 
                 797 
               
               
                 42 
                 42 
                 126 
                 210 
                 294 
                 378 
                 462 
                 546 
                 630 
                 714 
                 798 
               
               
                 43 
                 43 
                 127 
                 211 
                 295 
                 379 
                 463 
                 547 
                 631 
                 715 
                 799 
               
               
                 44 
                 44 
                 128 
                 212 
                 296 
                 380 
                 464 
                 548 
                 632 
                 716 
                 800 
               
               
                 45 
                 45 
                 129 
                 213 
                 297 
                 381 
                 465 
                 549 
                 633 
                 717 
                 801 
               
               
                 46 
                 46 
                 130 
                 214 
                 298 
                 382 
                 466 
                 550 
                 634 
                 718 
                 802 
               
               
                 47 
                 47 
                 131 
                 215 
                 299 
                 383 
                 467 
                 551 
                 635 
                 719 
                 803 
               
               
                 48 
                 48 
                 132 
                 216 
                 300 
                 384 
                 468 
                 552 
                 636 
                 720 
                 804 
               
               
                 49 
                 49 
                 133 
                 217 
                 301 
                 385 
                 469 
                 553 
                 637 
                 721 
                 805 
               
               
                 50 
                 50 
                 134 
                 218 
                 302 
                 386 
                 470 
                 554 
                 638 
                 722 
                 806 
               
               
                 51 
                 51 
                 135 
                 219 
                 303 
                 387 
                 471 
                 555 
                 639 
                 723 
                 807 
               
               
                 52 
                 52 
                 136 
                 220 
                 304 
                 388 
                 472 
                 556 
                 640 
                 724 
                 808 
               
               
                 53 
                 53 
                 137 
                 221 
                 305 
                 389 
                 473 
                 557 
                 641 
                 725 
                 809 
               
               
                 54 
                 54 
                 138 
                 222 
                 306 
                 390 
                 474 
                 558 
                 642 
                 726 
                 810 
               
               
                 55 
                 55 
                 139 
                 223 
                 307 
                 391 
                 475 
                 559 
                 643 
                 727 
                 811 
               
               
                 56 
                 56 
                 140 
                 224 
                 308 
                 392 
                 476 
                 560 
                 644 
                 728 
                 812 
               
               
                 57 
                 57 
                 141 
                 225 
                 309 
                 393 
                 477 
                 561 
                 645 
                 729 
                 813 
               
               
                 58 
                 58 
                 142 
                 226 
                 310 
                 394 
                 478 
                 562 
                 646 
                 730 
                 814 
               
               
                 59 
                 59 
                 143 
                 227 
                 311 
                 395 
                 479 
                 563 
                 647 
                 731 
                 815 
               
               
                 60 
                 60 
                 144 
                 228 
                 312 
                 396 
                 480 
                 564 
                 648 
                 732 
                 816 
               
               
                 61 
                 61 
                 145 
                 229 
                 313 
                 397 
                 481 
                 565 
                 649 
                 733 
                 817 
               
               
                 62 
                 62 
                 146 
                 230 
                 314 
                 398 
                 482 
                 566 
                 650 
                 734 
                 818 
               
               
                 63 
                 63 
                 147 
                 231 
                 315 
                 399 
                 483 
                 567 
                 651 
                 735 
                 819 
               
               
                 64 
                 64 
                 148 
                 232 
                 316 
                 400 
                 484 
                 568 
                 652 
                 736 
                 820 
               
               
                 65 
                 65 
                 149 
                 233 
                 317 
                 401 
                 485 
                 569 
                 653 
                 737 
                 821 
               
               
                 66 
                 66 
                 150 
                 234 
                 318 
                 402 
                 486 
                 570 
                 654 
                 738 
                 822 
               
               
                 67 
                 67 
                 151 
                 235 
                 319 
                 403 
                 487 
                 571 
                 655 
                 739 
                 823 
               
               
                 68 
                 68 
                 152 
                 236 
                 320 
                 404 
                 488 
                 572 
                 656 
                 740 
                 824 
               
               
                 69 
                 69 
                 153 
                 237 
                 321 
                 405 
                 489 
                 573 
                 657 
                 741 
                 825 
               
               
                 70 
                 70 
                 154 
                 238 
                 322 
                 406 
                 490 
                 574 
                 658 
                 742 
                 826 
               
               
                 71 
                 71 
                 155 
                 239 
                 323 
                 407 
                 491 
                 575 
                 659 
                 743 
                 827 
               
               
                 72 
                 72 
                 156 
                 240 
                 324 
                 408 
                 492 
                 576 
                 660 
                 744 
                 828 
               
               
                 73 
                 73 
                 157 
                 241 
                 325 
                 409 
                 493 
                 577 
                 661 
                 745 
                 829 
               
               
                 74 
                 74 
                 158 
                 242 
                 326 
                 410 
                 494 
                 578 
                 662 
                 746 
                 830 
               
               
                 75 
                 75 
                 159 
                 243 
                 327 
                 411 
                 495 
                 579 
                 663 
                 747 
                 831 
               
               
                 76 
                 76 
                 160 
                 244 
                 328 
                 412 
                 496 
                 580 
                 664 
                 748 
                 832 
               
               
                 77 
                 77 
                 161 
                 245 
                 329 
                 413 
                 497 
                 581 
                 665 
                 749 
                 833 
               
               
                 78 
                 78 
                 162 
                 246 
                 330 
                 414 
                 498 
                 582 
                 666 
                 750 
                 834 
               
               
                 79 
                 79 
                 163 
                 247 
                 331 
                 415 
                 499 
                 583 
                 667 
                 751 
                 835 
               
               
                 80 
                 80 
                 164 
                 248 
                 332 
                 416 
                 500 
                 584 
                 668 
                 752 
                 836 
               
               
                 81 
                 81 
                 165 
                 249 
                 333 
                 417 
                 501 
                 585 
                 669 
                 753 
                 837 
               
               
                 82 
                 82 
                 166 
                 250 
                 334 
                 418 
                 502 
                 586 
                 670 
                 754 
                 838 
               
               
                 83 
                 83 
                 167 
                 251 
                 335 
                 419 
                 503 
                 587 
                 671 
                 755 
                 839 
               
               
                 84 
                 84 
                 168 
                 252 
                 336 
                 420 
                 504 
                 588 
                 672 
                 756 
                 840 
               
               
                 85 
                 25 
                 26 
                 27 
                 28 
                 29 
                 30 
                 31 
                 32 
                 33 
                 34 
                 35 
                 36 
                 37 
                 38 
                 39 
                 40 
               
               
                 86 
                 113 
                 114 
                 115 
                 116 
                 117 
                 118 
                 119 
                 120 
                 121 
                 122 
                 123 
                 124 
                 125 
                 126 
                 127 
                 128 
               
               
                 87 
                 201 
                 202 
                 203 
                 204 
                 205 
                 206 
                 207 
                 208 
                 209 
                 210 
                 211 
                 212 
                 213 
                 214 
                 215 
                 216 
               
               
                 88 
                 289 
                 290 
                 291 
                 292 
                 293 
                 294 
                 295 
                 296 
                 297 
                 298 
                 299 
                 300 
                 301 
                 302 
                 303 
                 304 
               
               
                 89 
                 377 
                 378 
                 379 
                 380 
                 381 
                 382 
                 383 
                 384 
                 385 
                 386 
                 387 
                 388 
                 389 
                 390 
                 391 
                 392 
               
               
                 90 
                 465 
                 466 
                 467 
                 468 
                 469 
                 470 
                 471 
                 472 
                 473 
                 474 
                 475 
                 476 
                 477 
                 478 
                 479 
                 480 
               
               
                 91 
                 553 
                 554 
                 555 
                 556 
                 557 
                 558 
                 559 
                 560 
                 561 
                 562 
                 563 
                 564 
                 565 
                 566 
                 567 
                 568 
               
               
                 92 
                 641 
                 642 
                 643 
                 644 
                 645 
                 646 
                 647 
                 648 
                 649 
                 650 
                 651 
                 652 
                 653 
                 654 
                 655 
                 656 
               
               
                 93 
                 729 
                 730 
                 731 
                 732 
                 733 
                 734 
                 735 
                 736 
                 737 
                 738 
                 739 
                 740 
                 741 
                 742 
                 743 
                 744 
               
               
                 94 
                 817 
                 818 
                 819 
                 820 
                 821 
                 822 
                 823 
                 824 
                 825 
                 826 
                 827 
                 828 
                 829 
                 830 
                 831 
                 832 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 10 
               
               
                   
               
               
                 Example 5 screening pool design. 
               
               
                 55 seq 4 &amp; SP screening pool design 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Screening 
                   
               
               
                 pool # 
                 Unique pools contained in each screening pool 
               
               
                   
               
             
          
           
               
                  1 1 
                 1 
                 46 
                 91 
                 136 
                 181 
                 226 
                 271 
                 316 
                 361 
                 406 
               
               
                  2 
                 2 
                 47 
                 92 
                 137 
                 182 
                 227 
                 272 
                 317 
                 362 
                 407 
               
               
                  3 
                 3 
                 48 
                 93 
                 138 
                 183 
                 228 
                 273 
                 318 
                 363 
                 408 
               
               
                  4 
                 4 
                 49 
                 94 
                 139 
                 184 
                 229 
                 274 
                 319 
                 364 
                 409 
               
               
                  5 
                 5 
                 50 
                 95 
                 140 
                 185 
                 230 
                 275 
                 320 
                 365 
                 410 
               
               
                  6 
                 6 
                 51 
                 96 
                 141 
                 186 
                 231 
                 276 
                 321 
                 366 
                 411 
               
               
                  7 
                 7 
                 52 
                 97 
                 142 
                 187 
                 232 
                 277 
                 322 
                 367 
                 412 
               
               
                  8 
                 8 
                 53 
                 98 
                 143 
                 188 
                 233 
                 278 
                 323 
                 368 
                 413 
               
               
                  9 
                 9 
                 54 
                 99 
                 144 
                 189 
                 234 
                 279 
                 324 
                 369 
                 414 
               
               
                 10 
                 10 
                 55 
                 100 
                 145 
                 190 
                 235 
                 280 
                 325 
                 370 
                 415 
               
               
                 11 
                 11 
                 56 
                 101 
                 146 
                 191 
                 236 
                 281 
                 326 
                 371 
                 416 
               
               
                 12 
                 12 
                 57 
                 102 
                 147 
                 192 
                 237 
                 282 
                 327 
                 372 
                 417 
               
               
                 13 
                 13 
                 58 
                 103 
                 148 
                 193 
                 238 
                 283 
                 328 
                 373 
                 418 
               
               
                 14 
                 14 
                 59 
                 104 
                 149 
                 194 
                 239 
                 284 
                 329 
                 374 
                 419 
               
               
                 15 
                 15 
                 60 
                 105 
                 150 
                 195 
                 240 
                 285 
                 330 
                 375 
                 420 
               
               
                 16 
                 16 
                 61 
                 106 
                 151 
                 196 
                 241 
                 286 
                 331 
                 376 
                 421 
               
               
                 17 
                 17 
                 62 
                 107 
                 152 
                 197 
                 242 
                 287 
                 332 
                 377 
                 422 
               
               
                 18 
                 18 
                 63 
                 108 
                 153 
                 198 
                 243 
                 288 
                 333 
                 378 
                 423 
               
               
                 19 
                 19 
                 64 
                 109 
                 154 
                 199 
                 244 
                 289 
                 334 
                 379 
                 424 
               
               
                 20 
                 20 
                 65 
                 110 
                 155 
                 200 
                 245 
                 290 
                 335 
                 380 
                 425 
               
               
                 21 
                 21 
                 66 
                 111 
                 156 
                 201 
                 246 
                 291 
                 336 
                 381 
                 426 
               
               
                 22 
                 22 
                 67 
                 112 
                 157 
                 202 
                 247 
                 292 
                 337 
                 382 
                 427 
               
               
                 23 
                 23 
                 68 
                 113 
                 158 
                 203 
                 248 
                 293 
                 338 
                 383 
                 428 
               
               
                 24 
                 24 
                 69 
                 114 
                 159 
                 204 
                 249 
                 294 
                 339 
                 384 
                 429 
               
               
                 25 
                 25 
                 70 
                 115 
                 160 
                 205 
                 250 
                 295 
                 340 
                 385 
                 430 
               
               
                 26 
                 26 
                 71 
                 116 
                 161 
                 206 
                 251 
                 296 
                 341 
                 386 
                 431 
               
               
                 27 
                 27 
                 72 
                 117 
                 162 
                 207 
                 252 
                 297 
                 342 
                 387 
                 432 
               
               
                 28 
                 28 
                 73 
                 118 
                 163 
                 208 
                 253 
                 298 
                 343 
                 388 
                 433 
               
               
                 29 
                 29 
                 74 
                 119 
                 164 
                 209 
                 254 
                 299 
                 344 
                 389 
                 434 
               
               
                 30 
                 30 
                 75 
                 120 
                 165 
                 210 
                 255 
                 300 
                 345 
                 390 
                 435 
               
               
                 31 
                 31 
                 76 
                 121 
                 166 
                 211 
                 256 
                 301 
                 346 
                 391 
                 436 
               
               
                 32 
                 32 
                 77 
                 122 
                 167 
                 212 
                 257 
                 302 
                 347 
                 392 
                 437 
               
               
                 33 
                 33 
                 78 
                 123 
                 168 
                 213 
                 258 
                 303 
                 348 
                 393 
                 438 
               
               
                 34 
                 34 
                 79 
                 124 
                 169 
                 214 
                 259 
                 304 
                 349 
                 394 
                 439 
               
               
                 35 
                 35 
                 80 
                 125 
                 170 
                 215 
                 260 
                 305 
                 350 
                 395 
                 440 
               
               
                 36 
                 36 
                 81 
                 126 
                 171 
                 216 
                 261 
                 306 
                 351 
                 396 
                 441 
               
               
                 37 
                 37 
                 82 
                 127 
                 172 
                 217 
                 262 
                 307 
                 352 
                 397 
                 442 
               
               
                 38 
                 38 
                 83 
                 128 
                 173 
                 218 
                 263 
                 308 
                 353 
                 398 
                 443 
               
               
                 39 
                 39 
                 84 
                 129 
                 174 
                 219 
                 264 
                 309 
                 354 
                 399 
                 444 
               
               
                 40 
                 40 
                 85 
                 130 
                 175 
                 220 
                 265 
                 310 
                 355 
                 400 
                 445 
               
               
                 41 
                 41 
                 86 
                 131 
                 176 
                 221 
                 266 
                 311 
                 356 
                 401 
                 446 
               
               
                 42 
                 42 
                 87 
                 132 
                 177 
                 222 
                 267 
                 312 
                 357 
                 402 
                 447 
               
               
                 43 
                 43 
                 88 
                 133 
                 178 
                 223 
                 268 
                 313 
                 358 
                 403 
                 448 
               
               
                 44 
                 44 
                 89 
                 134 
                 179 
                 224 
                 269 
                 314 
                 359 
                 404 
                 449 
               
               
                 45 
                 45 
                 90 
                 135 
                 180 
                 225 
                 270 
                 315 
                 360 
                 405 
                 450 
               
               
                 46 
                 25 
                 26 
                 27 
                 28 
                 29 
                 30 
                 31 
                 32 
                 33 
                 34 
               
               
                 47 
                 113 
                 114 
                 115 
                 116 
                 117 
                 118 
                 119 
                 120 
                 121 
                 122 
               
               
                 48 
                 201 
                 202 
                 203 
                 204 
                 205 
                 206 
                 207 
                 208 
                 209 
                 210 
               
               
                 49 
                 289 
                 290 
                 291 
                 292 
                 293 
                 294 
                 295 
                 296 
                 297 
                 298 
               
               
                 50 
                 377 
                 378 
                 379 
                 380 
                 381 
                 382 
                 383 
                 384 
                 385 
                 386 
               
               
                 51 
                 465 
                 466 
                 467 
                 468 
                 469 
                 470 
                 471 
                 472 
                 473 
                 474 
               
               
                 52 
                 553 
                 554 
                 555 
                 556 
                 557 
                 558 
                 559 
                 560 
                 561 
                 562 
               
               
                 53 
                 641 
                 642 
                 643 
                 644 
                 645 
                 646 
                 647 
                 648 
                 649 
                 650 
               
               
                 54 
                 729 
                 730 
                 731 
                 732 
                 733 
                 734 
                 735 
                 736 
                 737 
                 738 
               
               
                 55 
                 817 
                 818 
                 819 
                 820 
                 821 
                 822 
                 823 
                 824 
                 825 
                 826 
               
               
                   
               
             
          
           
               
                 Screening 
                   
               
               
                 pool # 
                 Unique pools contained in each screening pool 
               
               
                   
               
             
          
           
               
                  1 
                 451 
                 496 
                 541 
                 586 
                 631 
                 676 
                 721 
                 766 
                 811 
                 856 
               
               
                  2 
                 452 
                 497 
                 542 
                 587 
                 632 
                 677 
                 722 
                 767 
                 812 
                 857 
               
               
                  3 
                 453 
                 498 
                 543 
                 588 
                 633 
                 678 
                 723 
                 768 
                 813 
                 858 
               
               
                  4 
                 454 
                 499 
                 544 
                 589 
                 634 
                 679 
                 724 
                 769 
                 814 
                 859 
               
               
                  5 
                 455 
                 500 
                 545 
                 590 
                 635 
                 680 
                 725 
                 770 
                 815 
                 860 
               
               
                  6 
                 456 
                 501 
                 546 
                 591 
                 636 
                 681 
                 726 
                 771 
                 816 
                 861 
               
               
                  7 
                 457 
                 502 
                 547 
                 592 
                 637 
                 682 
                 727 
                 772 
                 817 
                 862 
               
               
                  8 
                 458 
                 503 
                 548 
                 593 
                 638 
                 683 
                 728 
                 773 
                 818 
                 863 
               
               
                  9 
                 459 
                 504 
                 549 
                 594 
                 639 
                 684 
                 729 
                 774 
                 819 
                 864 
               
               
                 10 
                 460 
                 505 
                 550 
                 595 
                 640 
                 685 
                 730 
                 775 
                 820 
                 865 
               
               
                 11 
                 461 
                 506 
                 551 
                 596 
                 641 
                 686 
                 731 
                 776 
                 821 
                 866 
               
               
                 12 
                 462 
                 507 
                 552 
                 597 
                 642 
                 687 
                 732 
                 777 
                 822 
                 867 
               
               
                 13 
                 463 
                 508 
                 553 
                 598 
                 643 
                 688 
                 733 
                 778 
                 823 
                 868 
               
               
                 14 
                 464 
                 509 
                 554 
                 599 
                 644 
                 689 
                 734 
                 779 
                 824 
                 869 
               
               
                 15 
                 465 
                 510 
                 555 
                 600 
                 645 
                 690 
                 735 
                 780 
                 825 
                 870 
               
               
                 16 
                 466 
                 511 
                 556 
                 601 
                 646 
                 691 
                 736 
                 781 
                 826 
                 871 
               
               
                 17 
                 467 
                 512 
                 557 
                 602 
                 647 
                 692 
                 737 
                 782 
                 827 
                 872 
               
               
                 18 
                 468 
                 513 
                 558 
                 603 
                 648 
                 693 
                 738 
                 783 
                 828 
                 873 
               
               
                 19 
                 469 
                 514 
                 559 
                 604 
                 649 
                 694 
                 739 
                 784 
                 829 
                 874 
               
               
                 20 
                 470 
                 515 
                 560 
                 605 
                 650 
                 695 
                 740 
                 785 
                 830 
                 875 
               
               
                 21 
                 471 
                 516 
                 561 
                 606 
                 651 
                 696 
                 741 
                 786 
                 831 
                 876 
               
               
                 22 
                 472 
                 517 
                 562 
                 607 
                 652 
                 697 
                 742 
                 787 
                 832 
                 877 
               
               
                 23 
                 473 
                 518 
                 563 
                 608 
                 653 
                 698 
                 743 
                 788 
                 833 
                 878 
               
               
                 24 
                 474 
                 519 
                 564 
                 609 
                 654 
                 699 
                 744 
                 789 
                 834 
                 879 
               
               
                 25 
                 475 
                 520 
                 565 
                 610 
                 655 
                 700 
                 745 
                 790 
                 835 
                 880 
               
               
                 26 
                 476 
                 521 
                 566 
                 611 
                 656 
                 701 
                 746 
                 791 
                 836 
               
               
                 27 
                 477 
                 522 
                 567 
                 612 
                 657 
                 702 
                 747 
                 792 
                 837 
               
               
                 28 
                 478 
                 523 
                 568 
                 613 
                 658 
                 703 
                 748 
                 793 
                 838 
               
               
                 29 
                 479 
                 524 
                 569 
                 614 
                 659 
                 704 
                 749 
                 794 
                 839 
               
               
                 30 
                 480 
                 525 
                 570 
                 615 
                 660 
                 705 
                 750 
                 795 
                 840 
               
               
                 31 
                 481 
                 526 
                 571 
                 616 
                 661 
                 706 
                 751 
                 796 
                 841 
               
               
                 32 
                 482 
                 527 
                 572 
                 617 
                 662 
                 707 
                 752 
                 797 
                 842 
               
               
                 33 
                 483 
                 528 
                 573 
                 618 
                 663 
                 708 
                 753 
                 798 
                 843 
               
               
                 34 
                 484 
                 529 
                 574 
                 619 
                 664 
                 709 
                 754 
                 799 
                 844 
               
               
                 35 
                 485 
                 530 
                 575 
                 620 
                 665 
                 710 
                 755 
                 800 
                 845 
               
               
                 36 
                 486 
                 531 
                 576 
                 621 
                 666 
                 711 
                 756 
                 801 
                 846 
               
               
                 37 
                 487 
                 532 
                 577 
                 622 
                 667 
                 712 
                 757 
                 802 
                 847 
               
               
                 38 
                 488 
                 533 
                 578 
                 623 
                 668 
                 713 
                 758 
                 803 
                 848 
               
               
                 39 
                 489 
                 534 
                 579 
                 624 
                 669 
                 714 
                 759 
                 804 
                 849 
               
               
                 40 
                 490 
                 535 
                 580 
                 625 
                 670 
                 715 
                 760 
                 805 
                 850 
               
               
                 41 
                 491 
                 536 
                 581 
                 626 
                 671 
                 716 
                 761 
                 806 
                 851 
               
               
                 42 
                 492 
                 537 
                 582 
                 627 
                 672 
                 717 
                 762 
                 807 
                 852 
               
               
                 43 
                 493 
                 538 
                 583 
                 628 
                 673 
                 718 
                 763 
                 808 
                 853 
               
               
                 44 
                 494 
                 539 
                 584 
                 629 
                 674 
                 719 
                 764 
                 809 
                 854 
               
               
                 45 
                 495 
                 540 
                 585 
                 630 
                 675 
                 720 
                 765 
                 810 
                 855 
               
               
                 46 
                 35 
                 36 
                 37 
                 38 
                 39 
                 40 
               
               
                 47 
                 123 
                 124 
                 125 
                 126 
                 127 
                 128 
               
               
                 48 
                 211 
                 212 
                 213 
                 214 
                 215 
                 216 
               
               
                 49 
                 299 
                 300 
                 301 
                 302 
                 303 
                 304 
               
               
                 50 
                 387 
                 388 
                 389 
                 390 
                 391 
                 392 
               
               
                 51 
                 475 
                 476 
                 477 
                 478 
                 479 
                 480 
               
               
                 52 
                 563 
                 564 
                 565 
                 566 
                 567 
                 568 
               
               
                 53 
                 651 
                 652 
                 653 
                 654 
                 655 
                 656 
               
               
                 54 
                 739 
                 740 
                 741 
                 742 
                 743 
                 744 
               
               
                 55 
                 827 
                 828 
                 829 
                 830 
                 831 
                 832 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 11 
               
               
                   
               
               
                 Example 5 screening pool design. 
               
               
                 45 seq 5 screening pool design 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Screening 
                   
               
               
                 pool # 
                 Unique pools contained in each screening pool 
               
               
                   
               
             
          
           
               
                 1 
                 1 
                 46 
                 91 
                 136 
                 181 
                 226 
                 271 
                 316 
                 361 
                 406 
                 451 
                 496 
                 541 
               
               
                  2 
                 2 
                 47 
                 92 
                 137 
                 182 
                 227 
                 272 
                 317 
                 362 
                 407 
                 452 
                 497 
                 542 
               
               
                  3 
                 3 
                 48 
                 93 
                 138 
                 183 
                 228 
                 273 
                 318 
                 363 
                 408 
                 453 
                 498 
                 543 
               
               
                  4 
                 4 
                 49 
                 94 
                 139 
                 184 
                 229 
                 274 
                 319 
                 364 
                 409 
                 454 
                 499 
                 544 
               
               
                  5 
                 5 
                 50 
                 95 
                 140 
                 185 
                 230 
                 275 
                 320 
                 365 
                 410 
                 455 
                 500 
                 545 
               
               
                  6 
                 6 
                 51 
                 96 
                 141 
                 186 
                 231 
                 276 
                 321 
                 366 
                 411 
                 456 
                 501 
                 546 
               
               
                  7 
                 7 
                 52 
                 97 
                 142 
                 187 
                 232 
                 277 
                 322 
                 367 
                 412 
                 457 
                 502 
                 547 
               
               
                  8 
                 8 
                 53 
                 98 
                 143 
                 188 
                 233 
                 278 
                 323 
                 368 
                 413 
                 458 
                 503 
                 548 
               
               
                  9 
                 9 
                 54 
                 99 
                 144 
                 189 
                 234 
                 279 
                 324 
                 369 
                 414 
                 459 
                 504 
                 549 
               
               
                 10 
                 10 
                 55 
                 100 
                 145 
                 190 
                 235 
                 280 
                 325 
                 370 
                 415 
                 460 
                 505 
                 550 
               
               
                 11 
                 11 
                 56 
                 101 
                 146 
                 191 
                 236 
                 281 
                 326 
                 371 
                 416 
                 461 
                 506 
                 551 
               
               
                 12 
                 12 
                 57 
                 102 
                 147 
                 192 
                 237 
                 282 
                 327 
                 372 
                 417 
                 462 
                 507 
                 552 
               
               
                 13 
                 13 
                 58 
                 103 
                 148 
                 193 
                 238 
                 283 
                 328 
                 373 
                 418 
                 463 
                 508 
                 553 
               
               
                 14 
                 14 
                 59 
                 104 
                 149 
                 194 
                 239 
                 284 
                 329 
                 374 
                 419 
                 464 
                 509 
                 554 
               
               
                 15 
                 15 
                 60 
                 105 
                 150 
                 195 
                 240 
                 285 
                 330 
                 375 
                 420 
                 465 
                 510 
                 555 
               
               
                 16 
                 16 
                 61 
                 106 
                 151 
                 196 
                 241 
                 286 
                 331 
                 376 
                 421 
                 466 
                 511 
                 556 
               
               
                 17 
                 17 
                 62 
                 107 
                 152 
                 197 
                 242 
                 287 
                 332 
                 377 
                 422 
                 467 
                 512 
                 557 
               
               
                 18 
                 18 
                 63 
                 108 
                 153 
                 198 
                 243 
                 288 
                 333 
                 378 
                 423 
                 468 
                 513 
                 558 
               
               
                 19 
                 19 
                 64 
                 109 
                 154 
                 199 
                 244 
                 289 
                 334 
                 379 
                 424 
                 469 
                 514 
                 559 
               
               
                 20 
                 20 
                 65 
                 110 
                 155 
                 200 
                 245 
                 290 
                 335 
                 380 
                 425 
                 470 
                 515 
                 560 
               
               
                 21 
                 21 
                 66 
                 111 
                 156 
                 201 
                 246 
                 291 
                 336 
                 381 
                 426 
                 471 
                 516 
                 561 
               
               
                 22 
                 22 
                 67 
                 112 
                 157 
                 202 
                 247 
                 292 
                 337 
                 382 
                 427 
                 472 
                 517 
                 562 
               
               
                 23 
                 23 
                 68 
                 113 
                 158 
                 203 
                 248 
                 293 
                 338 
                 383 
                 428 
                 473 
                 518 
                 563 
               
               
                 24 
                 24 
                 69 
                 114 
                 159 
                 204 
                 249 
                 294 
                 339 
                 384 
                 429 
                 474 
                 519 
                 564 
               
               
                 25 
                 25 
                 70 
                 115 
                 160 
                 205 
                 250 
                 295 
                 340 
                 385 
                 430 
                 475 
                 520 
                 565 
               
               
                 26 
                 26 
                 71 
                 116 
                 161 
                 206 
                 251 
                 296 
                 341 
                 386 
                 431 
                 476 
                 521 
                 566 
               
               
                 27 
                 27 
                 72 
                 117 
                 162 
                 207 
                 252 
                 297 
                 342 
                 387 
                 432 
                 477 
                 522 
                 567 
               
               
                 28 
                 28 
                 73 
                 118 
                 163 
                 208 
                 253 
                 298 
                 343 
                 388 
                 433 
                 478 
                 523 
                 568 
               
               
                 29 
                 29 
                 74 
                 119 
                 164 
                 209 
                 254 
                 299 
                 344 
                 389 
                 434 
                 479 
                 524 
                 569 
               
               
                 30 
                 30 
                 75 
                 120 
                 165 
                 210 
                 255 
                 300 
                 345 
                 390 
                 435 
                 480 
                 525 
                 570 
               
               
                 31 
                 31 
                 76 
                 121 
                 166 
                 211 
                 256 
                 301 
                 346 
                 391 
                 436 
                 481 
                 526 
                 571 
               
               
                 32 
                 32 
                 77 
                 122 
                 167 
                 212 
                 257 
                 302 
                 347 
                 392 
                 437 
                 482 
                 527 
                 572 
               
               
                 33 
                 33 
                 78 
                 123 
                 168 
                 213 
                 258 
                 303 
                 348 
                 393 
                 438 
                 483 
                 528 
                 573 
               
               
                 34 
                 34 
                 79 
                 124 
                 169 
                 214 
                 259 
                 304 
                 349 
                 394 
                 439 
                 484 
                 529 
                 574 
               
               
                 35 
                 35 
                 80 
                 125 
                 170 
                 215 
                 260 
                 305 
                 350 
                 395 
                 440 
                 485 
                 530 
                 575 
               
               
                 36 
                 36 
                 81 
                 126 
                 171 
                 216 
                 261 
                 306 
                 351 
                 396 
                 441 
                 486 
                 531 
                 576 
               
               
                 37 
                 37 
                 82 
                 127 
                 172 
                 217 
                 262 
                 307 
                 352 
                 397 
                 442 
                 487 
                 532 
                 577 
               
               
                 38 
                 38 
                 83 
                 128 
                 173 
                 218 
                 263 
                 308 
                 353 
                 398 
                 443 
                 488 
                 533 
                 578 
               
               
                 39 
                 39 
                 84 
                 129 
                 174 
                 219 
                 264 
                 309 
                 354 
                 399 
                 444 
                 489 
                 534 
                 579 
               
               
                 40 
                 40 
                 85 
                 130 
                 175 
                 220 
                 265 
                 310 
                 355 
                 400 
                 445 
                 490 
                 535 
                 580 
               
               
                 41 
                 41 
                 86 
                 131 
                 176 
                 221 
                 266 
                 311 
                 356 
                 401 
                 446 
                 491 
                 536 
                 581 
               
               
                 42 
                 42 
                 87 
                 132 
                 177 
                 222 
                 267 
                 312 
                 357 
                 402 
                 447 
                 492 
                 537 
                 582 
               
               
                 43 
                 43 
                 88 
                 133 
                 178 
                 223 
                 268 
                 313 
                 358 
                 403 
                 448 
                 493 
                 538 
                 583 
               
               
                 44 
                 44 
                 89 
                 134 
                 179 
                 224 
                 269 
                 314 
                 359 
                 404 
                 449 
                 494 
                 539 
                 584 
               
               
                 45 
                 45 
                 90 
                 135 
                 180 
                 225 
                 270 
                 315 
                 360 
                 405 
                 450 
                 495 
                 540 
                 585 
               
               
                   
               
             
          
           
               
                 Screening 
                   
               
               
                 pool # 
                 Unique pools contained in each screening pool 
               
               
                   
               
             
          
           
               
                  1 
                 586 
                 631 
                 676 
                 721 
                 766 
                 811 
                 856 
                 21 
                 194 
                 367 
                 540 
                 713 
               
               
                  2 
                 587 
                 632 
                 677 
                 722 
                 767 
                 812 
                 857 
                 22 
                 195 
                 368 
                 541 
                 714 
               
               
                  3 
                 588 
                 633 
                 678 
                 723 
                 768 
                 813 
                 858 
                 23 
                 196 
                 369 
                 542 
                 715 
               
               
                  4 
                 589 
                 634 
                 679 
                 724 
                 769 
                 814 
                 859 
                 24 
                 197 
                 370 
                 543 
                 716 
               
               
                  5 
                 590 
                 635 
                 680 
                 725 
                 770 
                 815 
                 860 
                 89 
                 198 
                 371 
                 544 
                 717 
               
               
                  6 
                 591 
                 636 
                 681 
                 726 
                 771 
                 816 
                 861 
                 90 
                 199 
                 372 
                 545 
                 718 
               
               
                  7 
                 592 
                 637 
                 682 
                 727 
                 772 
                 817 
                 862 
                 91 
                 200 
                 373 
                 546 
                 719 
               
               
                  8 
                 593 
                 638 
                 683 
                 728 
                 773 
                 818 
                 863 
                 92 
                 265 
                 374 
                 547 
                 720 
               
               
                  9 
                 594 
                 639 
                 684 
                 729 
                 774 
                 819 
                 864 
                 93 
                 266 
                 375 
                 548 
                 721 
               
               
                 10 
                 595 
                 640 
                 685 
                 730 
                 775 
                 820 
                 865 
                 94 
                 267 
                 376 
                 549 
                 722 
               
               
                 11 
                 596 
                 641 
                 686 
                 731 
                 776 
                 821 
                 866 
                 95 
                 268 
                 441 
                 550 
                 723 
               
               
                 12 
                 597 
                 642 
                 687 
                 732 
                 777 
                 822 
                 867 
                 96 
                 269 
                 442 
                 551 
                 724 
               
               
                 13 
                 598 
                 643 
                 688 
                 733 
                 778 
                 823 
                 868 
                 97 
                 270 
                 443 
                 552 
                 725 
               
               
                 14 
                 599 
                 644 
                 689 
                 734 
                 779 
                 824 
                 869 
                 98 
                 271 
                 444 
                 617 
                 726 
               
               
                 15 
                 600 
                 645 
                 690 
                 735 
                 780 
                 825 
                 870 
                 99 
                 272 
                 445 
                 618 
                 727 
               
               
                 16 
                 601 
                 646 
                 691 
                 736 
                 781 
                 826 
                 871 
                 100 
                 273 
                 446 
                 619 
                 728 
               
               
                 17 
                 602 
                 647 
                 692 
                 737 
                 782 
                 827 
                 872 
                 101 
                 274 
                 447 
                 620 
                 793 
               
               
                 18 
                 603 
                 648 
                 693 
                 738 
                 783 
                 828 
                 873 
                 102 
                 275 
                 448 
                 621 
                 794 
               
               
                 19 
                 604 
                 649 
                 694 
                 739 
                 784 
                 829 
                 874 
                 103 
                 276 
                 449 
                 622 
                 795 
               
               
                 20 
                 605 
                 650 
                 695 
                 740 
                 785 
                 830 
                 875 
                 104 
                 277 
                 450 
                 623 
                 796 
               
               
                 21 
                 606 
                 651 
                 696 
                 741 
                 786 
                 831 
                 876 
                 105 
                 278 
                 451 
                 624 
                 797 
               
               
                 22 
                 607 
                 652 
                 697 
                 742 
                 787 
                 832 
                 877 
                 106 
                 279 
                 452 
                 625 
                 798 
               
               
                 23 
                 608 
                 653 
                 698 
                 743 
                 788 
                 833 
                 878 
                 107 
                 280 
                 453 
                 626 
                 799 
               
               
                 24 
                 609 
                 654 
                 699 
                 744 
                 789 
                 834 
                 879 
                 108 
                 281 
                 454 
                 627 
                 800 
               
               
                 25 
                 610 
                 655 
                 700 
                 745 
                 790 
                 835 
                 880 
                 109 
                 282 
                 455 
                 628 
                 801 
               
               
                 26 
                 611 
                 656 
                 701 
                 746 
                 791 
                 836 
                 1 
                 110 
                 283 
                 456 
                 629 
                 802 
               
               
                 27 
                 612 
                 657 
                 702 
                 747 
                 792 
                 837 
                 2 
                 111 
                 284 
                 457 
                 630 
                 803 
               
               
                 28 
                 613 
                 658 
                 703 
                 748 
                 793 
                 838 
                 3 
                 112 
                 285 
                 458 
                 631 
                 804 
               
               
                 29 
                 614 
                 659 
                 704 
                 749 
                 794 
                 839 
                 4 
                 177 
                 286 
                 459 
                 632 
                 805 
               
               
                 30 
                 615 
                 660 
                 705 
                 750 
                 795 
                 840 
                 5 
                 178 
                 287 
                 460 
                 633 
                 806 
               
               
                 31 
                 616 
                 661 
                 706 
                 751 
                 796 
                 841 
                 6 
                 179 
                 288 
                 461 
                 634 
                 807 
               
               
                 32 
                 617 
                 662 
                 707 
                 752 
                 797 
                 842 
                 7 
                 180 
                 353 
                 462 
                 635 
                 808 
               
               
                 33 
                 618 
                 663 
                 708 
                 753 
                 798 
                 843 
                 8 
                 181 
                 354 
                 463 
                 636 
                 809 
               
               
                 34 
                 619 
                 664 
                 709 
                 754 
                 799 
                 844 
                 9 
                 182 
                 355 
                 464 
                 637 
                 810 
               
               
                 35 
                 620 
                 665 
                 710 
                 755 
                 800 
                 845 
                 10 
                 183 
                 356 
                 529 
                 638 
                 811 
               
               
                 36 
                 621 
                 666 
                 711 
                 756 
                 801 
                 846 
                 11 
                 184 
                 357 
                 530 
                 639 
                 812 
               
               
                 37 
                 622 
                 667 
                 712 
                 757 
                 802 
                 847 
                 12 
                 185 
                 358 
                 531 
                 640 
                 813 
               
               
                 38 
                 623 
                 668 
                 713 
                 758 
                 803 
                 848 
                 13 
                 186 
                 359 
                 532 
                 705 
                 814 
               
               
                 39 
                 624 
                 669 
                 714 
                 759 
                 804 
                 849 
                 14 
                 187 
                 360 
                 533 
                 706 
                 815 
               
               
                 40 
                 625 
                 670 
                 715 
                 760 
                 805 
                 850 
                 15 
                 188 
                 361 
                 534 
                 707 
                 816 
               
               
                 41 
                 626 
                 671 
                 716 
                 761 
                 806 
                 851 
                 16 
                 189 
                 362 
                 535 
                 708 
               
               
                 42 
                 627 
                 672 
                 717 
                 762 
                 807 
                 852 
                 17 
                 190 
                 363 
                 536 
                 709 
               
               
                 43 
                 628 
                 673 
                 718 
                 763 
                 808 
                 853 
                 18 
                 191 
                 364 
                 537 
                 710 
               
               
                 44 
                 629 
                 674 
                 719 
                 764 
                 809 
                 854 
                 19 
                 192 
                 365 
                 538 
                 711 
               
               
                 45 
                 630 
                 675 
                 720 
                 765 
                 810 
                 855 
                 20 
                 193 
                 366 
                 539 
                 712 
               
               
                   
               
             
          
         
       
     
     Tables 8, 9, 10 and 11 shows four of the many specific repooling designs that were tested to demonstrate the utility of this patent. 
     Tables 12-16 are data showing multiple embodiments of various randomization schemes for pooling a quantification of data loaded into the Matrix Pool Plate ( FIG. 5 ). 
     
       
         
               
             
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 12 
               
             
             
               
                   
               
               
                 Summary of various screening pool 
               
               
                 design unique clone identification. 
               
             
          
           
               
                 Pooling Summary with each clone 
                   
               
               
                 contained in 4 to 8 unique pools. 
               
             
          
           
               
                   
                   
                 Total 
                   
                   
               
               
                   
                   
                 possible 
               
               
                 Screening 
                   
                 instances 
                 Unique clone identification 
               
             
          
           
               
                 Pool size 
                 design 
                 of clone 
                 maximum 
                 −1 
                 −2 
                 −3 
               
               
                   
               
             
          
           
               
                 30 
                 rnd 4 
                 4 
                 86.4% 
                 13.0% 
                 0.6% 
                 0.0% 
               
               
                 30 
                 seq 4 
                 4 
                 83.7% 
                 16.0% 
                 0.3% 
                 0.0% 
               
               
                 45 
                 rnd 4 
                 4 
                 88.0% 
                 11.6% 
                 0.3% 
                 0.0% 
               
               
                 45 
                 seq 5 
                 5 
                 85.1% 
                 14.3% 
                 0.6% 
                 0.0% 
               
               
                 55 
                 seq 4 &amp; SP 
                 5 
                 91.2% 
                 8.5% 
                 0.2% 
                 0.0% 
               
               
                 61 
                 rnd 4 
                 4 
                 91.9% 
                 8.0% 
                 0.1% 
                 0.0% 
               
               
                 61 
                 seq 4 
                 4 
                 95.1% 
                 4.9% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 seq &amp; step 8 
                 8 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 seq 8 
                 8 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 seq &amp; rnd 8 
                 8 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 seq 6 
                 6 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 step 5 
                 5 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 seq 5 
                 5 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 89 
                 rnd 4 
                 4 
                 94.6% 
                 5.3% 
                 0.1% 
                 0.0% 
               
               
                 89 
                 seq 4 
                 4 
                 100.0% 
                 0.0% 
                 0.0% 
                 0.0% 
               
               
                 94 
                 seq 4 &amp; SP 
                 5 
                 99.3% 
                 0.7% 
                 0.0% 
                 0.0% 
               
               
                 94 
                 seq 5 
                 5 
                 96.8% 
                 3.2% 
                 0.0% 
                 0.0% 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 13 
               
             
             
               
                   
               
               
                 Summary of various screening pool 
               
               
                 design unique clone identification. 
               
             
          
           
               
                   
                 Possibilities to find one random clone 
                   
               
             
          
           
               
                 Screening 
                   
                 False positives found during identification 
                   
               
             
          
           
               
                 Pool size 
                 design 
                 &lt;9 
                 9–7 
                 7–5 
                 5–3 
                 2 
                 1 
                 0 
                 −1 
               
               
                   
               
             
          
           
               
                 30 
                 rnd 4 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 30 
                 seq 4 
               
               
                 45 
                 rnd 4 
               
               
                 45 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 4% 
                 3% 
                 46% 
                 48% 
                 4% 
               
               
                 55 
                 seq 4 &amp; 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 43% 
                 49% 
                 8% 
               
               
                   
                 SP 
               
               
                 61 
                 rnd 4 
               
               
                 61 
                 seq 4 
               
               
                 89 
                 seq &amp; step 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                   
                 8 
               
               
                 89 
                 seq 8 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 seq &amp; rnd 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                   
                 8 
               
               
                 89 
                 seq 6 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 step 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 rnd 4 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 95% 
                 5% 
               
               
                 89 
                 seq 4 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 94 
                 seq 4 &amp; 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                   
                 SP 
               
               
                 94 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 96% 
                 4% 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 14 
               
             
             
               
                   
               
               
                 Summary of various screening pool designs searching for one unique 
               
               
                 clone identification. 
               
             
          
           
               
                   
                 Possibilities to find one random clone 
                   
               
             
          
           
               
                 Screening 
                   
                 False positives found during identification 
                   
               
             
          
           
               
                 Pool size 
                 design 
                 &lt;9 
                 9–7 
                 7–5 
                 5–3 
                 2 
                 1 
                 0 
                 −1 
               
               
                   
               
             
          
           
               
                 30 
                 rnd 4 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 30 
                 seq 4 
               
               
                 45 
                 rnd 4 
               
               
                 45 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 4% 
                 3% 
                 46% 
                 48% 
                 4% 
               
               
                 55 
                 seq 4 &amp; 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 43% 
                 49% 
                 8% 
               
               
                   
                 SP 
               
               
                 61 
                 rnd 4 
               
               
                 61 
                 seq 4 
               
               
                 89 
                 seq &amp; step 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                   
                 8 
               
               
                 89 
                 seq 8 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 seq &amp; rnd 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                   
                 8 
               
               
                 89 
                 seq 6 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 step 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 89 
                 rnd 4 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 95% 
                 5% 
               
               
                 89 
                 seq 4 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                 94 
                 seq 4 &amp; 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 100% 
               
               
                   
                 SP 
               
               
                 94 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 96% 
                 4% 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 15 
               
             
             
               
                   
               
               
                 Summary of various screening pool designs 
               
               
                 searching for two unique clone identifications. 
               
               
                 Possibilities to find random sets of 2 unique 
               
               
                 but similar marker containing clones 
               
             
          
           
               
                   
                 False positives found during identification 
                   
               
             
          
           
               
                 Screening pool 
                 design 
                 6+ 
                 5 
                 4 
                 3 
                 2 
                 1 
                 0 
                 −1 
               
               
                   
               
             
          
           
               
                 30 
                 rnd 4 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 30 
                 seq 4 
               
               
                 45 
                 rnd 4 
               
               
                 45 
                 seq 5 
                 39% 
                 12% 
                 11% 
                 11% 
                 8% 
                 10% 
                 7% 
                 1% 
               
               
                 55 
                 seq 4 &amp; SP 
                 15% 
                 10% 
                 14% 
                 24% 
                 10% 
                 16% 
                 8% 
                 1% 
               
               
                 61 
                 rnd 4 
               
               
                 61 
                 seq 4 
               
               
                 89 
                 seq &amp; step 8 
                 4% 
                 4% 
                 4% 
                 6% 
                 22% 
                 33% 
                 21% 
                 0% 
               
               
                 89 
                 seq 8 
               
               
                 89 
                 seq &amp; rnd 8 
                 0% 
                 0% 
                 0% 
                 0% 
                 2% 
                 5% 
                 61% 
                 29% 
               
               
                 89 
                 seq 6 
               
               
                 89 
                 step 5 
                 1% 
                 1% 
                 3% 
                 11% 
                 14% 
                 41% 
                 29% 
                 0% 
               
               
                 89 
                 seq 5 
                 0% 
                 0% 
                 1% 
                 1% 
                 7% 
                 22% 
                 63% 
                 6% 
               
               
                 89 
                 rnd 4 
                 0% 
                 0% 
                 1% 
                 4% 
                 10% 
                 20% 
                 64% 
                 1% 
               
               
                 89 
                 seq 4 
                 1% 
                 1% 
                 4% 
                 5% 
                 17% 
                 38% 
                 34% 
                 0% 
               
               
                 94 
                 seq 4 &amp; SP 
                 0% 
                 0% 
                 0% 
                 0% 
                 5% 
                 11% 
                 84% 
                 0% 
               
               
                 94 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 1% 
                 7% 
                 20% 
                 69% 
                 3% 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 16 
               
             
             
               
                   
               
               
                 Summary of various screening pool designs searching for three unique 
               
               
                 clone identifications. 
               
             
          
           
               
                 Possibilities to find random sets of 3 
                   
               
               
                 unique but similar marker containing clones 
               
             
          
           
               
                   
                   
                 False positives found 
                   
               
               
                 Screening 
                   
                 during identification 
               
             
          
           
               
                 Pool size 
                 design 
                 &gt;15 
                 14 
                 13 
                 12 
                 11 
                 10 
                 9 
                 8 
                 7 
                 6 
                 5 
                 4 
                 3 
                 2 
                 1 
                 0 
                 −1 
                 −2 
               
               
                   
               
             
          
           
               
                 30 
                 rnd 4 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 30 
                 seq 4 
               
               
                 45 
                 rnd 4 
               
               
                 45 
                 seq 5 
                 89% 
                 0% 
                 1% 
                 1% 
                 0% 
                 2% 
                 0% 
                 2% 
                 0% 
                 2% 
                 0% 
                 0% 
                 1% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
               
               
                 55 
                 seg 4 
                 61% 
                 8% 
                 3% 
                 2% 
                 2% 
                 5% 
                 7% 
                 1% 
                 0% 
                 3% 
                 3% 
                 0% 
                 1% 
                 1% 
                 0% 
                 0% 
                 0% 
                 0% 
               
               
                   
                 &amp; SP 
               
               
                 61 
                 rnd 4 
               
               
                 61 
                 seq 4 
               
               
                 89 
                 seg &amp; 
                 20% 
                 4% 
                 3% 
                 4% 
                 3% 
                 6% 
                 9% 
                 10% 
                 13% 
                 8% 
                 9% 
                 6% 
                 3% 
                 1% 
                 0% 
                 1% 
                 0% 
                 0% 
               
               
                   
                 step 8 
               
               
                 89 
                 seq 8 
                 17% 
                 4% 
                 3% 
                 4% 
                 14% 
                 6% 
                 7% 
                 6% 
                 10% 
                 14% 
                 8% 
                 10% 
                 5% 
                 0% 
                 1% 
                 0% 
                 1% 
                 0% 
               
               
                 89 
                 seq &amp; 
                 2% 
                 2% 
                 5% 
                 5% 
                 3% 
                 2% 
                 11% 
                 9% 
                 13% 
                 14% 
                 13% 
                 10% 
                 6% 
                 2% 
                 2% 
                 1% 
                 0% 
                 0% 
               
               
                   
                 rnd 8 
               
               
                 89 
                 seq 6 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 2% 
                 3% 
                 2% 
                 7% 
                 19% 
                 20% 
                 27% 
                 17% 
                 1% 
               
               
                 89 
                 step 5 
                 2% 
                 2% 
                 5% 
                 5% 
                 3% 
                 2% 
                 11% 
                 9% 
                 13% 
                 14% 
                 13% 
                 10% 
                 6% 
                 2% 
                 2% 
                 1% 
                 0% 
                 0% 
               
               
                 89 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 1% 
                 5% 
                 3% 
                 7% 
                 14% 
                 14% 
                 16% 
                 28% 
                 9% 
                 3% 
                 0% 
               
               
                 89 
                 rnd 4 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 5% 
                 5% 
                 9% 
                 8% 
                 13% 
                 17% 
                 17% 
                 19% 
                 7% 
                 0% 
                 0% 
               
               
                 89 
                 seq 4 
                 2% 
                 2% 
                 1% 
                 3% 
                 2% 
                 2% 
                 10% 
                 17% 
                 14% 
                 19% 
                 8% 
                 10% 
                 8% 
                 1% 
                 0% 
                 1% 
                 0% 
                 0% 
               
               
                 94 
                 seq 4 &amp; 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 1% 
                 0% 
                 1% 
                 8% 
                 14% 
                 14% 
                 24% 
                 32% 
                 8% 
                 2% 
                 0% 
               
               
                   
                 SP 
               
               
                 94 
                 seq 5 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 0% 
                 2% 
                 2% 
                 0% 
                 6% 
                 15% 
                 37% 
                 26% 
                 11% 
                 1% 
                 0% 
               
               
                   
               
             
          
         
       
     
     Tables 13, 14, 15 and 16 show data collected form various pooling designs. 
     In order to facilitate quick and accurate analysis of user screening data, we have developed a computer program which identifies the appropriate plate and well position of all potential positive clones. The results will be processed with error correction algorithms to enhance the reliability of the results and compensate for false negative data and false positive data (inherent in many screening technologies like PCR). The results will be displayed as probability scores indicating the likelihood of the resulting plate and well position being correct. 
     While the invention has been described with reference to more than one preferred embodiment, it is to be clearly understood by those skilled in the art that the invention is not limited thereto to these two embodiments. The general concept of separating the large library set into multiple superpools and then making one, or more than one, set(s) of matrix pools formed by re-pooling a subset of the unique pools into screening pools that will be screened. Each unique pool can be placed in 0, 1 or more than one screening pools, depending on the redundancy of identification required.