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).

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
       [0001]    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  
         [0002]    Not Applicable  
         REFERENCE TO SEQUENCE LISTING, A TABLE, OR A COMPUTER PROGRAM LISTING COMPACT DISK APPENDIX  
         [0003]    Not Applicable  
           [0004]    Field of Search: 341/., 435/., 435/ . . . , 435/DIG21, 435/DIG51, 536/23.1, 700/., 707/101  
           [0005]    References Cited [Referenced By]  
           [0006]    U.S. Patent Documents  
           [0007]    5,780,222  
           [0008]    6,126074  
           [0009]    6,477,699  
           [0010]    6,582,923  
           [0011]    6,607,888  
           [0012]    6,665,829  
           [0013]    6,706,867  
           [0014]    6,727,068  
           [0015]    6,727,071  
           [0016]    Other References  
           [0017]    Ausubel et al., “Short Protocols in Molecular Biology”, Wiley and Sons, New York.  
           [0018]    Sambrook et al., “Molecular Cloning, A Laboratory Manual”, Cold Springs Harbor Press, New York.  
           [0019]    Torney et al., “Pooling of a Total Genomic BAC Library”, US DOE Contract W-7405-ENG-36.  
           [0020]    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  
         [0021]    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.  
           [0022]    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).  
         BREIF SUMMARY OF THE INVENTION  
         [0023]    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.  
           [0024]    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 patterncan be used to for quality assurance and for assisting in deconvolution if desired.  
         BREIF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS  
         [0025]    Not Applicable 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0026]    EXAMPLE 1  
         [0027]    Detailed Description of Pools &amp; Superpools:  
         [0028]    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.  
         [0029]    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).  
         [0030]    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.  
         [0031]    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:  
         [0032]    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                    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                      
 
         [0033]    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.                    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                      
 
         [0034]    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.                    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                      
 
         [0035]    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.            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                  
 
         [0036]    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.                    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                      
 
         [0037]    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.  
         [0038]    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 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).  
         [0039]    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  
       [0040]    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).  
         [0041]    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).  
         [0042]    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.  
         [0043]    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.  
         [0044]    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.  
         [0045]    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.  
         [0046]    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; . . . ; 8A10,12B11,12C12 . . . 12P1        4   1A4,1B5,1C6 . . . 1P19; 2A5,2B6,2C7 . . . 2P20; . . . ; 8A11,12B12,12C13 . . . 12P2        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                  
 
         [0047]    [0047]                                                                                                                     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                    
         [0048]    [0048]                                                                               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                    
         [0049]    [0049]                                                                                               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                    
         [0050]    [0050]                                                                                                                               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                    
         [0051]    [0051]                                                                                                                                                   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                    
         [0052]    Tables 8, 9, 10 and 11 shows four of the many specific repooling designs that were tested to demonstrate the utility of this patent.  
                                                                                                           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%                  
 
         [0053]    [0053]                                                                                                                                   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%                    
         [0054]    [0054]                                                                                                                                   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%                    
         [0055]    [0055]                                                                                                                 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%                    
         [0056]    [0056]                                                                                                                                                                                                               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%                    
         [0057]    Tables 13, 14, 15 and 16 show data collected form various pooling designs.  
         [0058]    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.  
         [0059]    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.