Patent Publication Number: US-2006014186-A1

Title: Methods for genotype screening of a strain disposed on an adsorbent carrier

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS  
      This application claims priority under 35 U.S.C. §120 as a CONTINUATION-IN-PART APPLICATION of a co-pending application entitled “System, Method and Apparatus for Transgenic and Targeted Mutagenesis Screening” which was filed on Sep. 4, 2001, and was assigned U.S. application Ser. No. 09/945,952 (the “&#39;952 Application”), U.S. patent application Ser. No. 11/074,995 filed Mar. 8, 2005, and U.S. patent application Ser. No. ______ filed Jun. 24, 2005, entitled “Methods for Genotype Screening” the entire disclosures of which are incorporated herein by reference for all that it teaches. This application and the &#39;952 Application also claim priority under 35 U.S.C. §119(e), based on U.S. Provisional Application Ser. No. 60/230,371, filed Sep. 6, 2000, the entire disclosure of which is incorporated herein by reference for all that it teaches. 
    
    
     FIELD OF THE INVENTION  
      This invention relates to methods for genotype screening. More specifically, this invention relates to various methods to detect or screen for at least one designated genetic sequences in a plurality of biological samples, disposed on an adsorbent carrier.  
     BACKGROUND OF THE INVENTION  
      Genomic modification resulting from mutations in the DNA of an organism can be transferred to the progeny if such mutations are present in the gametes of the organism, referred to as germ-line mutations. These mutations may arise from genetic manipulation of the DNA using recombinant DNA technology or may be introduced by challenging the DNA by chemical or physical means. DNA introduced via recombinant DNA technology can be derived from many sources, including but not limited to DNA from viruses, mycoplasm, bacteria, fungi, yeast, and chordates including mammals such as humans.  
      Recombinant DNA technology allows for the introduction, deletion or replacement of DNA of an organism. Random introduction of DNA into a cell can be achieved by technologies such as transfection (including electroporation, lipofection), injection (pronuclear injection, nuclear transplantation) or transduction (viral infection). Random mutations (point mutations, deletions, amplifications) can be generated by treatment of cells with chemical mutagens or submitting them to physical insult such as X-irradiation or linear energy transfer irradiation (LET). Targeted addition, deletion or replacement of DNA in an organism (either inducible or non-inducible) is achieved via homologous recombination. Inducible systems employ sequence-specific recombinases such as Cre-LoxP (U.S. Pat. Nos. 5,654,182 and 5,677,177) and FLP/FRT (U.S. Pat. No. 5,527,695).  
      Transgenic organisms are organisms that carry DNA sequences (be it genes or gene segments) derived from another or the same species, stably integrated randomly into their genome. Transgenic mammals are generally created by microinjection of DNA into the pronucleus of fertilized eggs, a technique in which the number of DNA copies or the integration site of the DNA into the host genome is uncontrollable. A transgenic line or strain refers to an organism that transmits the foreign DNA sequences to its offspring.  
      Genotype screening is used to determine if a genome possesses specific genetic sequences that exist endogenously or have been modified, mutated or genetically engineered. Genomic nucleic acid is screened for these modifications, mutations or endogenous conditions. Genomic nucleic acid is challenging to work with because of its size. The genomic nucleic acid includes both coding and noncoding regions. Therefore, the genomic nucleic acid contains exons and introns, promoter and gene regulation regions, telomeres, origins or replication and nonfunctional intergenic nucleic acid. The genomic nucleic acid is a double stranded molecule which is methylated. cDNA and PCR-amplicons differs in that the molecules are much smaller. Additionally, biochemical modification events, such as methylation, do not occur with the smaller molecules. Shena, M (2000) DNA Microarrays: A Practical Approach. Oxford University Press, New York, N.Y.  
      Genotype screening is currently done manually. The present manual system is time-consuming and can provide variable results depending on the laboratory and even depending on skill of laboratory workers. Presently, a researcher using Southern blot technology may require greater than a week to screen a tissue sample for a transgene or a targeted mutation.  
      In an alternative technology, up to thirty PCR (polymerase chain reaction) can be conducted in an Eppendorf microtube® (Brinkmann Instruments, Westbury, N.Y.) and separated on a gel. This process in most laboratories requires 3 to 7 days. A need exists in the industry to provide a system and method for more accurate, faster and high volume genotype screening.  
      Additionally, as researchers continue to use transgenic species in research specific information about the progeny of the transgenic species is of vital importance. An emerging technique in mouse mutant breeding is producing ‘homozygous’ transgenic conditions. During the initial creation of transgenic animals the transgene sequence integrates randomly into the host genome. Moreover, the number of transgene insertions also varies. Once the transgene is established in the genome, some investigators are interested in having this/these transgene(s) on the corresponding chromosome. The preferred mechanism for getting both chromosomes to have the transgene(s), is by breeding two transgenic animals from the same strain together. The goal is to identify homozygous animals that can then be bred to each other to ensure continual homozygous progeny. Typically, such transgenic animals are difficult to genotype by traditional PCR methods as accurate quantification is not possible with fragment-based analysis.  
     SUMMARY OF THE INVENTION  
      The present invention provides a unique solution to the above-described problems by providing a method for rapid genotype screening. In particular, this invention provides a method to rapidly report screening results to a remote user from a screening laboratory for a plurality of biological samples disposed on an adsorbent carrier. Efficient screening of a plurality of biological samples can be achieved by placing the sample to be screened in a well of a microwell container. The biological samples in the microwell containers are lysed to release at least a portion of intact genomic nucleic acid and cellular debris. In one embodiment, a standard concentration of purified genomic nucleic acid is obtained by saturating the binding ability of the magnetic particles and by regulating the amount of genomic nucleic acid released. The purified genomic nucleic acid are screened to obtain screening results. The screening results are reported to a remote user. These screening results can include information on whether a designated genetic sequence is present in an organism and the zygosity of designated genetic sequences. Additionally, the zygosity of a transgene can be quantitatively determined and reported to a remote user.  
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
      A more complete understanding of the invention and its advantages will be apparent from the following Description of the Preferred Embodiment(s) taken in conjunction with the accompanying drawings, wherein:  
       FIG. 1  is an illustrative overview of the remote automated testing procedures of the present invention.  
       FIG. 2  is a block diagram of one embodiment of the system.  
       FIG. 3  is a block diagram of the ordering procedure.  
       FIG. 4  is a block diagram of account registration.  
       FIGS. 5-6  illustrate the survey of work and sample identification sections.  
       FIG. 7A  is a block diagram of the laboratory process system.  
       FIG. 7B  is a block diagram of the laboratory process system.  
       FIG. 7C  is a block diagram of the laboratory process system.  
       FIG. 7D  is a block diagram of the laboratory process system.  
       FIG. 8  is a block diagram of standard laboratory stations.  
       FIG. 9  is a screen display illustrating a document on the transgenic screening laboratory  20 &#39;s web site relating to an outcome file.  
       FIG. 10  is a graphical representation of the results.  
       FIG. 11  is a graphical representation of signal magnitude.  
       FIG. 12  is a graphical representation of signal magnitude.  
       FIG. 13  is a graphical representation of signal magnitude.  
       FIGS. 14 and 15  illustrate a preferred device for performing the functions of a Lysing Station and an Automated Accessioning Station as described herein, including an oven ( FIG. 15 ) for incubating the samples.  
       FIG. 16  illustrates a preferred device for performing the functions of an Isolation/Purification Station as described herein.  
       FIG. 17  illustrates a preferred device for drying samples.  
       FIG. 18  illustrates a preferred device for performing the functions of a Screening Station as described herein.  
       FIG. 19  illustrates a preferred device for performing the functions of a Detection Station as described herein.  
       FIG. 20A  shows a schematic diagram of two swab holders.  
       FIG. 20B  shows a cross-sectional view of a swab holder.  
       FIG. 21  shows a schematic diagram of a kit.  
       FIGS. 22-25  show a representative screening result for human data. 
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
      The present invention provides a method for high volume genotype screening. This invention provides a method for rapid identification of an organism, whose genome possesses specific genetic sequences that exist endogenously or has been modified, mutated or genetically engineered. All patents, patent applications and articles discussed or referred to in this specification are hereby incorporated by reference.  
     1. DEFINITIONS  
      The following terms and acronyms are used throughout the detailed description.  
      Alox5-KO  
                              TGCCCAGCGGTCCTATCTAGAGGTCATTCTCTCCACAGAGCGAGTCAAGAACCACTG   (SEQ ID NO. 1)           GCAGGAAGACCTCATGTTTGGCTACCAGTTCCTGAATGGCTGCAACCCAGTAATTCT       ACCGGGTAGGGGAGGCGCTTTTCCCAAGGCAGTCTGGAGCATGCGCTTTAGCAGCC       CCGCTGGCACTTGGCGCTACACAAGTGGCCTCTGGCCTCGCACACATTCCACATCCA       CCGGTAGCGCCAACCGGCTCCGTTCTTTGGTGGCCCCTTCGCGCCACCTTCTACTCCT       CCCCTAGTCAGGAAGTTCCCCCCCGCCCCGCAGCTCGCGTCGTGCAGGACGTGACAA       ATGGAAGTAGCACGTCTCACTAGTCTCGTGCAGATGGACAGCACCGCTGAGCAATG       GAAGCGGGTAGGCCTTTGGGGCAGCGGCCAATAGCAGCTTTGCTCCTTCGCTTTCTG       GGCTCAGAGGCTGGGAAGGGGTGGGTCCGGGGGCGGGCTCAGGG          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer Seq.: 
                   
                   
               
               
                   
                 TTGGCTACCAGTTCCTGAATGG 
                 (SEQ ID NO. 2) 
               
               
                   
                   
               
               
                   
                 Reverse Primer Seq.: 
               
               
                   
                 CAGACTGCCTTGGGAAAAGC 
                 (SEQ ID NO. 3) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CTGCAACCCAGTAATTC 
                 (SEQ ID NO. 4) 
               
            
           
         
       
     
      Alox5-WT  
                              AAGAACCACTGGCAGGAAGACCTCATGTTTGGCTACCAGTTCCTGAATGGCTGCAAC   (SEQ ID NO. 5)           CCAGTACTCATCAAGCGCTGCACAGCGTTGCCCCCGAAGCTCCCAGTGACCACAGA       GATGGTGGAGTGCAGCCTAGAGCGGCAGCTCAGTTTAGAACA          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer Seq.: 
                   
                   
               
               
                   
                 TTGGCTACCAGTTCCTGAATGG 
                 (SEQ ID NO. 6) 
               
               
                   
                   
               
               
                   
                 Reverse Primer Seq.: 
               
               
                   
                 CTGTGGTCACTGGGAGCTT 
                 (SEQ ID NO. 7) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CTGCAACCCAGTACTCAT 
                 (SEQ ID NO. 8) 
               
            
           
         
       
     
      APC Min  
                          (SEQ ID NO. 9)                                 TATCATGTCTCCCGGCTCAAGTCTGCCATCCCTTCACGTTAGGAAACAGAAAGCTCT               AGAAGCTGAGCTAGATGCTCAGCATTTATCAGAAACCTTCGACAACATTGACAACCT           AAGTCCCAAGGCCTCTCACCGGAGTAAGCAGAGACACAAGCAGAATCTTTATGGTG           ACTATGCTTTTGACGCCAATCGACATGATGATAGTAGGTCAGACAATTTCAATACTG           GAAACATGACTGTTCTTTCACCATATTTAAATACTACGGTATTGCCCAGCTCTTCTTC           CTCAAGGGGAAGTTTAGACAGTTCTCGTTCTGAGAAAGACAGAAGTTAGGAGAGAG           AGCGAGGTATTGGCCTCAGTGCTTACCATCCAACAACAGAAAATGCAGGAACCTCA           TCAAAACGAGGTCTGCAGATCACTACCACTGCAGCCCAGATAGCCAAAGTTATGGA           AGAAGTATCAGCCATTCATACCTCCCAGGACGACAGAAGTTCTGCTTCTACCACCGA           GTTCCATTGTGTGGCAGACGACAGGAGTGCGGCACGAAGAAGCTCTGCCTNNNNNN           NNNNNNNNNNNNNNNNNNNCTTCACTAAGTCGGAAAATTCAAATAGGACATGCTCT           ATGCCTTATGCCAAAGTGGAATATAAACGATCTTCAAATGACAGTTTAAATA           GTGTCACTAGTA          
 
     
       
         
           
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
               
               
                   
                 GGGAAGTTTAGACAGTTCTCGTTCT (SEQ ID NO. 10) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 GTAAGCACTGAGGCCAATACCT (SEQ ID NO. 11) 
               
               
                   
                   
               
               
                   
                 Probe 1: 
               
               
                   
                 CTCTCTCCAAACTTC (SEQ ID NO. 12) 
               
               
                   
                   
               
               
                   
                 Probe 2: 
               
               
                   
                 TCTCTCTCCTAACTTC (SEQ ID NO. 13) 
               
            
           
         
       
     
      Bgal  
                          (SEQ ID NO. 14)                                 GTTGAGAATGAGTACGGGTCCTACTTTGCCTGCGATTACGACTACCTACGCTTCCTG               GTGCACCGCTTCCGCTACCATCTGGGTAATGACGTCATTCTCTTCACCACCGACGGA           GCAAGTGAAAAAATGCTGAAGTGTGGGACCCTGCAGGACCTGTACGCCACAGTGGA           TTTTGGAACAG          
 
     
       
         
           
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer Seq.: 
                   
               
               
                   
                 CACCGCTTCCGCTACCAT (SEQ ID NO. 15) 
               
               
                   
                   
               
               
                   
                 Reverse Primer Seq.: 
               
               
                   
                 GCTCCGTCGGTGGTGAAG (SEQ ID NO. 16) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CTGGGTAATGACGTCATTCT (SEQ ID NO. 17) 
               
            
           
         
       
     
      complementary—chemical affinity between nitrogenous bases as a result of hydrogen bonding. Responsible for the base pairing between nucleic acid strands. Klug, W. S. and Cummings, M. R. (1997) Concepts of Genetics, fifth ed., Prentice-Hall, Upper Saddle River, N.J.  
      copy number—the number of transgenes that have randomly integrated into the genome.  
      Cjun—(housekeeping or reference sequence)  
                          (SEQ ID NO. 18)                                 GACCGGTAACAAGTGGCCGGGAGCGAACTTTTGCAAATCTCTTCTGCGCCTTAAGGC               TGCCACCGAGACTGTAAAGAAAAGGGAGAAGAGGAACCTATACTCATACCAGTTCG           CACAGGCGGCTGAAGTTGGGCGAGCGCTAGCCGCGGCTGCCTAGCGTCCCCCTCCC           CCTCACAGCGGAGGAGGGGACAGTTGTCGGAGGCCGGGCGGCAGAGCCCGATCGC           GGGCTTCCACCGAGAATTCCGTGACGACTGGTCAGCACCGCCGGAGAGCCGCTGTT           GCTGGGACTGGTCTGCGGGCTCCAAGGAACCGCTGCTCCCCGAGAGCGCTCCGTGA           GTGACCGCGACTTTTCAAAGCTCGGCATCGCGCGGGAGCCTACCAACGTGAGTGCT           AGCGGAGTCTTAACCCTGCGCTCCCTGGAGCGAACTGGGGAGGAGGGCTCAGGGGG           AAGCACTGCCGTCTGGAGCGCACGCTCCTAAACAAACTTTGTTACAGAAGCGGGGA           CGCGCGGGTATCCCCCCGCTTCCCGGCGCGCTGTTGCGGCCCCGAAACTTCTGCGCA           CAGCCCAGGCTAACCCCGCGTGAAGTGACGGACCGTTCTATGACTGCAAAGATGGA           AACGACCTTCTACGACGATGCCCTCAACGCCTCGTTCCTCCAGTCCGAGAGCGGTGC           CTACGGCTACAGTAACCCTAAGATCCTAAAACAGAGCATGACCTTGAACCTGGCCG           ACCCGGTGGGCAGTCTGAAGCCGCACCTCCGCGCCAAGAACTCGGACCTTCTCACGT           CGCCCGACGTCGGGCTGCTCAAGCTGGCGTCGCCGGAGCTGGAGCGCCTGATCATC           CAGTCCAGCAATGGGCACATCACCACTACACCGACCCCCACCCAGTTCTTGTGCCCC           AAGAACGTGACCGACGAGCAGGAGGGCTTCGCCGAGGGCTTCGTGCGCGCCCTGGC           TGAACTGCATAGCCAGAACACGCTTCCCAGTGTCACCTCCGCGGCACAGCCGGTCA           GCGGGGCGGGCATGGTGGCTCCCGCGGTGGCCTCAGTAGCAGGCGCTGGCGGCGGT           GGTGGCTACAGCGCCAGCCTGCACAGTGAGCCTCCGGTCTACGCCAACCTCAGCAA           CTTCAACCCGGGTGCGCTGAGCAGCGGCGGTGGGGCGCCCTCCTATGGCGCGGCCG           GGCTGGCCTTTCCCTCGCAGCCGCAGCAGCAGCAGCAGCCGCCTCAGCCGCCGCAC           CACTTGCCCCAACAGATCCCGGTGCAGCACCCGCGGCTGCAAGCCCTGAAGGAAGA           GCCGCAGACCGTGCCGGAGATGCCGGGAGAGACGCCGCCCCTGTCCCCTATCGACA           TGGAGTCTCAGGAGCGGATCAAGGCAGAGAGGAAGCGCATGAGGAACCGCATTGCC           GCCTCCAAGTGCCGGAAAAGGAAGCTGGAGCGGATCGCTCGGCTAGAGGAAAAAGT           GAAAACCTTGAAAGCGCAAAACTCCGAGCTGGCATCCACGGCCAACATGCTCAGGG           AACAGGTGGCACAGCTTAAGCAGAAAGTCATGAACCACGTTAACAGTGGGTGCCAA           CTCATGCTAACGCAGCAGTTGCAAACGTTTTGAGAACAGACTGTCAGGGCTGAGGG           GCAATGGAAGAAAAAAAATAACAGAGACAAACTTGAGAACTTGACTGGTTGCGACA           GAGAAAAAAAAAGTGTCCGAGTACTGAAGCCAAGGGTACACAAGATGGACTGGGTT           GCGACCTGACGGCGCCCCCAGTGTGCTGGAGTGGGAAGGACGTGGCGCGCCTGGCT           TTGGCGTGGAGCCAGAGAGCAGCGGCCTATTGGCCGGCAGACTTTGCGGACGGGCT           GTGCCCGCGCGCGACCAGAACGATGGACTTTTCGTTAACATTGACCAAGAACTGCAT           GGACCTAACATTCGATCTCATTCAGTATTAAAGGGGGGTGGGAGGGGTTACAAACT           GCAATAGAGACTGTAGATTGCTTCTGTAGTGCTCCTTAACACAAAGCAGGGAGGGCT           GGGAAGGGGGGGGAGGCTTGTAAGTGCCAGGCTAGACTGCAGATGAACTCCCCTGG           CCTGCCTCTCTCAACTGTGTATGTACATATATATTTTTTTTTAATTTGATGAAAGCTG           ATTACTGTCAATAAACAGCTTCCTGCCTTTGTAAGTTATTCCATGTTTGTTTGTTTGG           GTGTCCTGCCC          
 
     
       
         
           
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
               
               
                   
                 GAGTGCTAGCGGAGTCTTAACC (SEQ ID NO. 19) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CTCCAGACGGCAGTGCTT (SEQ ID NO. 20) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 AAGCACTGCCGTCTGGAG (SEQ ID NO. 21) 
               
            
           
         
       
     
      Cre  
                          (SEQ ID: NO. 22)                         ATGCCCAAGAAGAAGAGGAAGGTGTCCAATTTACTGACCGTACACCAAAATTTGCC           TGCATTACCGGTCGATGCAACGAGTGATGAGGTTCGCAAGAACCTGATGGACATGTT       CAGGGATCGCCAGGCGTTTTCTGAGCATACCTGGAAAATGCTTCTGTCCGTTTGCCG       GTCGTGGGCGGCATGGTGCAAGTTGAATAACCGGAAATGGTTTCCCGCAGAACCTG       AAGATGTTCGCGATTATCTTCTATATCTTCAGGCGCGCGGTCTGGCAGTAAAAACTA       TCCAGCAACATTTGGGCCAGCTAAACATGCTTCATCGTCGGTCCGGGCTGCCACGAC       CAAGTGACAGCAATGCTGTTTCACTGGTTATGCGGCGGATCCGAAAAGAAAACGTT       GATGCCGGTGAACGTGCAAAACAGGCTCTAGCGTTCGAACGCACTGATTTCGACCA       GGTTCGTTCACTCATGGAAAATAGCGATCGCTGCCAGGATATACGTAATCTGGCATT       TCTGGGGATTGCTTATAACACCCTGTTACGTATAGCCGAAATTGCCAGGATCAGGGT       TAAAGATATCTCACGTACTGACGGTGGGAGAATGTTAATCCATATTGGCAGAACGA       AAACGCTGGTTAGCACCGCAGGTGTAGAGAAGGCACTTAGCCTGGGGGTAACTAAA       CTGGTCGAGCGATGGATTTCCGTCTCTGGTGTAGCTGATGATCCGAATAACTACCTG       TTTTGCCGGGTCAGAAAAAATGGTGTTGCCGCGCCATCTGCCACCAGCCAGCTATCA       ACTCGCGCCCTGGAAGGGATTTTTGAAGCAACTCATCGATTGATTTACGGCGCTAAG       GATGACTCTGGTCAGAGATACCTGGCCTGGTCTGGACACAGTGCCCGTGTCGGAGCC       GCGCGAGATATGGCCCGCGCTGGAGTTTCAATACCGGAGATCATGCAAGCTGGTGG       CTGGACCAATGTAAATATTGTCATGAACTATATCCGTAACCTGGATAGTGAAACAGG       GGCAATGGTGCGCCTGCTGGAAGATGGCGATTAGCCATTAACGCGTAAATGATTGCT       ATAATTATTTGATAT          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 TTAATCCATATTGGCAGAACGAAAACG 
                 (SEQ ID: NO. 23) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CAGGCTAAGTGCCTTCTCTACA 
                 (SEQ ID: NO. 24) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CCTGCGGTGCTAACC 
                 (SEQ ID: NO. 25) 
               
            
           
         
       
     
      designated genetic sequence—includes a transgenic insert, a selectable marker, microsatellite loci, recombinant site or any gene or gene segment.  
      DNA (deoxyribonucleic acid)—One of the two main types of nucleic acid, consisting of a long, unbranched macromolecule formed from one, or more commonly, two, strands of linked deoxyribonucleotides, the 3″-phosphate group of each constituent deoxyribonucleotide being joined in 3′,5′-phosphodiester linkage to the 5′-hydroxyl group of the deoxyribose moiety of the next one.  Oxford Dictionary of Biochemistry and Molecular Biology;  p. 182.  
      embryonic stem cells (ES cells)—a cell of the early embryo that can replicate indefinitely and which can differentiate into other cells; stem cells serve as a continuous source of new cells.  
      genome—all the genetic material in the chromosomes of a particular organism; its size is generally given as its total number of base pairs.  
      genomic nucleic acid—The genomic nucleic acid includes both coding and noncoding regions. Therefore, the genomic nucleic acid contains exons and introns, promoter and gene regulation regions, telomeres, origins or replication and nonfunctional intergenic nucleic acid. The genomic nucleic acid is a double stranded molecule which is methylated. cDNA and PCR-amplicons differs in that the molecules are much smaller. Additionally, biochemical modification events, such as methylation, do not occur with the smaller molecules. Shena, M (2000) DNA Microarrays: A Practical Approach. Oxford University Press, New York, N.Y.  
      genotype—genetic constitution of an individual cell or organism that can include at least one designated gene sequence.  
      hemizygous—a situation within a cell or organism where only one copy of a gene, group of genes or genetic sequence is present instead of two copies in a diploid genome.  
      heterozygosity—the state of having two different genes (alleles) at one or more corresponding loci on homologous chromosomes.  
      homozygosity—The state of having the same genes (alleles) at one or more corresponding homologous chromosomes.  
      HumanTTTy8  
                          (SEQ ID NO. 26)                         AAAGAAGAGCAGCACGTCATACCCAAGACCAACATCTCTCAGTGTTTCACGCTAAC           CCAAGGAGAGACACTAGCAGTCTTCTCTGCAGGACCCCTTGAATTTACATTGAATTC       CATCCCCAGCCGAGCAGGTGCTTAAAGTCAACAGGGGACACTCCATTTTCTTGGAAT       TTCATTCTGGCAAAGAGGGTGTGAGCAGCAATAAG          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer Seq.: 
                   
                   
               
               
                   
                 GCAGGACCCCTTGAATTTACATTGA 
                 (SEQ ID NO. 27) 
               
               
                   
                   
               
               
                   
                 Reverse Primer Seq.: 
               
               
                   
                 TGGAGTGTCCCCTGTTGACT 
                 (SEQ ID NO. 28) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CCGAGCAGGTGCTTAA 
                 (SEQ ID NO. 29) 
               
            
           
         
       
     
      Hygromycin  
                          (SEQ ID: No. 30)                         ATGAAAAAGCCTGAACTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTC           GACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTCAGC       TTCGATGTAGGAGGGCGTGGATATGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTC       TACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCCGCGCTCCCGATTCCGGAA       GTGCTTGACATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCCGTGCA       CAGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCG       GTCGCGGAGGCCATGGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTT       CGGCCCATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGATTTCATATG       CGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAG       TGCGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGA       AGTCCGGCACCTCGTGCACGCGGATTTCGGCTCCAACAATGTCCTGACGGACAATGG       CCGCATAACAGCGGTCATTGACTGGAGCGAGGCGATGTTCGGGGATTCCCAATACG       AGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGACGC       GCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTAT       ATGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGAT       GATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGTCCGATCCGGAGCCGGGAC       TGTCGGGCGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTG       TAGAAGTACTCGCCGATAGTGGAAACCGACGCCCCAGCACTCGTCCGAGGGCAAAG       GAATAG          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 CGCAAGGAATCGGTCAATACACTA 
                 (SEQ ID NO.: 31) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CACAGTTTGCCAGTGATACACATG 
                 (SEQ ID NO.: 32) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CATGGCGTGATTTCAT 
                 (SEQ ID NO.: 33) 
               
            
           
         
       
     
      internet—a collection of interconnected (public and/or private) networks that are linked together by a set of standard protocols to form a global, distributed network. The World Wide Web (hereinafter web) refers to both a distributed collection of interlinked, user viewable hypertext documents (commonly referred to as web pages) that are accessible via the Internet and the user and server software components which provide user access to such documents using standard Internet protocols.  
      line—A line is a group of organisms bred for a genotype (i.e. at least one designated genetic sequence).  
      MHV  
                              TATAAGAGTGATTGGCGTCCGTACGTACCCTCTCAACTCTAAAACTCTTGTAGTTTA   (SEQ ID NO.: 34)           AATCTAATCTAAACTTTATAAACGGCACTTCCTGCGTGTCCATGCCCGCGGGCCTGG       TCTTGTCATAGTGCTGACATTTGTAGTTCCTTGACTTTCGTTCTCTGCCAGTGACGTG       TCCATTCGGCGCCAGCAGCCCACCCATAGGTTGCATAATGGCAAAGATGGGCAAAT       ACGGTCTCGGCTTCAAATGGGCCCCAGAATTTCCATGGATGCTTCCGAACGCATCGG       AGAAGTTGGGTAACCCTGAGAGGTCAGAGGAGGATGGGTTTTGCCCCTCTGCTGCG       CAAGAACCGAAAGTTAAAGGAAAAACTTTGGTTAATCACGTGAGGGTGAATTGTAG       CCGGCTTCCAGCTTTGGAATGCTGTGTTCAGTCTGCCATAATCCGTGATATTTTTGTA       GATGAGGATCCCCAGAAGGTGGAGGCCTCAACTATGATGGCATTGCAGTTCGGTAG       TGCCGTCTTGGTTAAGCCATCCAAGCGCTTGTCTATTCAGGCATGGACTAATTTGGG       TGTGCTTCCCAAAACAGCTGCCATGGGGTTGTTCAAGCGCGTCTGCCTGTGTAACAC       CAGGGAGTGCTCTTGTGACGCCCACGTGGCCTTTCACCTTTTTACGGTCCAACCCGA       TGGTGTATGCCTGGGTAATGGCCGTTTTATAGGCTGGTTCGTTCCAGTCACAGCCAT       ACCGGAGTATGCGAAGCAGTGGTTGCAACCCTGGTCCATCCTTCTTCGTAAGGGTGG       TAACAAAGGGTCTGTGACATCCGGCCACTTCCGCCGCGCTGTTACCATGCCTGTGTA       TGACTTTAATGTAGAGGATGCTTGTGAGGAGGTTCATCTTAACCCGAAGGGTAAGTA       CTCCTGCAAGGCGTATGCTCTTCTTAAGGGCTATCGCGGTGTTAAGCCCATCCTGTTT       GTGGACCAGTATGGTTGCGACTATACTGGATGTCTCGCCAAGGGTCTTGAGGACTAT       GGCGATCTCACCTTGAGTGAGATGAAGGAGTTGTTCCCTGTGTGGCGTGACTCCTTG       GATAGTGAAGTCCTTGTGGCTTGGCACGTTGATCGAGATCCTCGGGCTGCTATGCGT       CTGCAGACTCTTGCTACTGTACGTTGCATTGATTATGTGGGCCAACCGACCGAGGAT       GTGGTGGATGGAGATGTGGTAGTGCGTGAGCCTGCTCATCTTCTCGCAGCCAATGCC       ATTGTTAAAAGACTCCCCCGTTTGGTGGAGACTATGCTGTATACGGATTCGTCCGTT       ACAGAATTCTGTTATAAAACCAAGCTGTGTGAATGCGGTTTTATCACGCAGTTTGGC       TATGTGGATTGTTGTGGTGACACCTGCGATTTTCGTGGGTGGGTTGCCGGCAATATG       ATGGATGGCTTTCCATGTCCAGGGTGTACCAAAAATTATATGCCCTGGGAATTGGAG       GCCCAGTCATCAGGTGTTATACCAGAAGGAGGTGTTCTATTCACTCAGAGCACTGAT       ACAGTGAATCGTGAGTCCTTTAAGCTCTACGGTCATGCTGTTGTGCCTTTTGGTTCTG       CTGTGTATTGGAGCCCTTGCCCAGGTATGTGGCTTCCAGTAATTTGGTCTTCTGTTAA       GTCATACTCTGGTTTGACTTATACAGGAGTAGTTGGTTGTAAGGCAATTGTTCAAGA       GACAGACGCTATATGTCGTTCTCTGTATATGGATTATGTCCAGCACAAGTGTGGCAA       TCTCGAGCAGAGAGCTATCCTTGGATTGGACGATGTCTATCATAGACAGTTGCTTGT       GAATAGGGGTGACTATAGTCTCCTCCTTGAGAATGTGGATTTGTTTGTTAAGCGGCG       CGCTGAATTTGCTTGCAAATTCGCCACCTGTGGAGATGGTCTTGTACCCCTCCTACTA       GATGGTTTAGTGCCCCGCAGTTATTATTTGATTAAGAGTGGTCAAGCTTTCACCTCTA       TGATGGTTAATTTTAGCCATGAGGTGACTGACATGTGTATGGACATGGCTTTATTGTT       CATGCATGATGTTAAAGTGGCCACTAAGTATGTTAAGAAGGTTACTGGCAAACTGGC       CGTGCGCTTTAAAGCGTTGGGTGTAGCCGTTGTCAGAAAAATTACTGAATGGTTTGA       TTTAGCCGTGGACATTGCTGCTAGTGCCGCTGGATGGCTTTGCTACCAGCTGGTAAA       TGGCTTATTTGCAGTGGCCAATGGTGTTATAACCTTTGTACAGGAGGTGCCTGAGCT       TGTCAAGAATTTTGTTGACAAGTTCAAGGCATTTTTCAAGGTTTTGATCGACTCTATG       TCGGTTTCTATCTTGTCTGGACTTACTGTTGTCAAGACTGCCTCAAATAGGGTGTGTC       TTGCTGGCAGTAAGGTTTATGAAGTTGTGCAGAAATCTTTGTCTGCATATGTTATGCC       TGTGGGTTGCAGTGAAGCCACTTGTTTGGTGGGTGAGATTGAACCTGCAGTTTTTGA       AGATGATGTTGTTGATGTGGTTAAAGCCCCATTAACATATCAAGGCTGTTGTAAGCC       ACCCACTTCTTTCGAGAAGATTTGTATTGTGGATAAATTGTATATGGCCAAGTGTGG       TGATCAATTTTACCCTGTGGTTGTTGATAACGACACTGTTGGCGTGTTAGATCAGTGC       TGGAGGTTTCCCTGTGCGGGCAAGAAAGTCGAGTTTAACGACAAGCCCAAAGTCAG       GAAGATACCCTCCACCCGTAAGATTAAGATCACCTTCGCACTGGATGCGACCTTTGA       TAGTGTTCTTTCGAAGGCGTGTTCAGAGTTTGAAGTTGATAAAGATGTTACATTGGA       TGAGCTGCTTGATGTTGTGCTTGACGCAGTTGAGAGTACGCTCAGCCCTTGTAAGGA       GCATGATGTGATAGGCACAAAAGTTTGTGCTTTACTTGATAGGTTGGCAGGAGATTA       TGTCTATCTTTTTGATGAGGGAGGCGATGAAGTGATCGCCCCGAGGATGTATTGTTC       CTTTTCTGCTCCTGATGATGAAGACTGCGTTGCAGCGGATGTTGTAGATGCAGATGA       AAACCAAGATGATGATGCTGAAGACTCAGCAGTCCTTGTCGCTGATACCCAAGAAG       AGGACGGCGTTGCCAAGGGGCAGGTTGAGGCGGATTCGGAAATTTGCGTTGCGCAT       ACTGGTAGTCAAGAAGAATTGGCTGAGCCTGATGCTGTCGGATCTCAAACTCCCATC       GCCTCTGCTGAGGAAACCGAAGTCGGAGAGGCAAGCGACAGGGAAGGGATTGCTG       AGGCGAAGGCAACTGTGTGTGCTGATGCTGTAGATGCCTGCCCCGATCAAGTGGAG       GCATTTGAAATTGAAAAGGTTGAAGACTCTATCTTGGATGAGCTTCAAACTGAACTT       AATGCGCCAGCGGACAAGACCTATGAGGATGTCTTGGCATTCGATGCCGTATGCTCA       GAGGCGTTGTCTGCATTCTATGCTGTGCCGAGTGATGAGACGCACTTTAAAGTGTGT       GGATTCTATTCGCCTGCTATAGAGCGCACTAATTGTTGGCTGCGTTCTACTTTGATAG       TAATGCAGAGTCTACCTTTGGAATTTAAAGACTTGGAGATGCAAAAGCTCTGGTTGT       CTTACAAGGCCGGCTATGACCAATGCTTTGTGGACAAACTAGTTAAGAGCGTGCCCA       AGTCTATTATCCTTCCACAAGGTGGTTATGTGGCAGATTTTGCCTATTTCTTTCTAAG       CCAGTGTAGCTTTAAAGCTTATGCTAACTGGCGTTGTTTAGAGTGTGACATGGAGTT       AAAGCTTCAAGGCTTGGACGCCATGTTTTTCTATGGGGACGTTGTGTCTCATATGTG       CAAGTGTGGTAATAGCATGACCTTGTTGTCTGCAGATATACCCTACACTTTGCATTTT       GGAGTGCGAGATGATAAGTTTTGCGCTTTTTACACGCCAAGAAAGGTCTTTAGGGCT       GCTTGTGCGGTAGATGTTAATGATTGTCACTCTATGGCTGTAGTAGAGGGCAAGCAA       ATTGATGGTAAAGTGGTTACCAAATTTATTGGTGACAAATTTGATTTTATGGTGGGT       TACGGGATGACATTTAGTATGTCTCCTTTTGAACTCGCCCAGTTATATGGTTCATGTA       TAACACCAAATGTTTGTTTTGTTAAAGGAGATGTTATAAAGGTTGTTCGCTTAGTTA       ATGCTGAAGTCATTGTTAACCCTGCTAATGGGCGTATGGCTCATGGTGCAGGTGTTG       CAGGTGCTATAGCTGAAAAGGCGGGCAGTGCTTTTATTAAAGAAACCTCCGATATG       GTGAAGGCTCAGGGCGTTTGCCAGGTTGGTGAATGCTATGAATCTGCCGGTGGTAAG       TTATGTAAAAAGGTGCTTAACATTGTAGGGCCAGATGCGCGAGGGCATGGCAAGCA       ATGCTATTCACTTTTAGAGCGTGCTTATCAGCATATTAATAAGTGTGACAATGTTGTC       ACTACTTTAATTTCGGCTGGTATATTTAGTGTGCCTACTGATGTCTCCCTAACTTACT       TACTTGGTGTAGTGACAAAGAATGTCATTCTTGTCAGTAACAACCAGGATGATTTTG       ATGTGATAGAGAAGTGTCAGGTGACCTCCGTTGCTGGTACCAAAGCGCTATCACTTC       AATTGGCCAAAAATTTGTGCCGTGATGTAAAGTTTGTGACGAATGCATGTAGTTCGC       TTTTTAGTGAATCTTGCTTTGTCTCAAGCTATGATGTGTTGCAGGAAGTTGAAGCGCT       GCGACATGATATACAATTGGATGATGATGCTCGTGTCTTTGTGCAGGCTAATATGGA       CTGTCTGCCCACAGACTGGCGTCTCGTTAACAAATTTGATAGTGTTGATGGTGTTAG       AACCATTAAGTATTTTGAATGCCCGGGCGGGATTTTTGTATCCAGCCAGGGCAAAAA       GTTTGGTTATGTTCAGAATGGTTCATTTAAGGAGGCGAGTGTTAGCCAAATAAGGGC       TTTACTCGCTAATAAGGTTGATGTCTTGTGTACTGTTGATGGTGTTAACTTCCGCTCC       TGCTGCGTAGCAGAGGGTGAAGTTTTTGGCAAGACATTAGGTTCAGTCTTTTGTGAT       GGCATAAATGTCACCAAAGTTAGGTGTAGTGCCATTTACAAGGGTAAGGTTTTCTTT       CAGTACAGTGATTTGTCCGAGGCAGATCTTGTGGCTGTTAAAGATGCCTTTGGTTTT       GATGAACCACAACTGCTGAAGTACTACACTATGCTTGGCATGTGTAAGTGGTCAGTA       GTTGTTTGTGGCAATTATTTTGCTTTCAAGCAGTCAAATAATAATTGCTATATAAATG       TGGCATGTTTAATGCTGCAACACTTGAGTTTAAAGTTTCCTAAGTGGCAATGGCAAG       AGGCTTGGAACGAGTTCCGCTCTGGTAAACCACTAAGGTTTGTGTCCTTGGTATTAG       CAAAGGGCAGCTTTAAATTTAATGAACCTTCTGATTCTATCGATTTTATGCGTGTGGT       GCTACGTGAAGCAGATTTGAGTGGTGCCACGTGCAATTTGGAATTTGTTTGTAAATG       TGGTGTGAAGCAAGAGCAGCGCAAAGGTGTTGACGCTGTTATGCATTTTGGTACGTT       GGATAAAGGTGATCTTGTCAGGGGTTATAATATCGCATGTACGTGCGGTAGTAAACT       TGTGCATTGCACCCAATTTAACGTACCATTTTTAATTTGCTCCAACACACCAGAGGG       TAGGAAACTGCCCGACGATGTTGTTGCAGCTAATATTTTTACTGGTGGTAGTGTGGG       CCATTACACGCATGTGAAATGTAAACCCAAGTACCAGCTTTATGATGCTTGTAATGT       TAATAAGGTTTCGGAGGCTAAGGGTAATTTTACCGATTGCCTCTACCTTAAAAATTT       AAAGCAAACTTTTTCGTCTGTGCTGACGACTTTTTATTTAGATGATGTAAAGTGTGTG       GAGTATAAGCCAGATTTATCGCAGTATTACTGTGAGTCTGGTAAATATTATACAAAA       CCCATTATTAAGGCCCAATTTAGAACATTTGAGAAGGTTGATGGTGTCTATACCAAC       TTTAAATTGGTGGGACATAGTATTGCTGAAAAACTCAATGCTAAGCTGGGATTTGAT       TGTAATTCTCCCTTTGTGGAGTATAAAATTACAGAGTGGCCAACAGCTACTGGAGAT       GTGGTGTTGGCTAGTGATGATTTGTATGTAAGTCGGTACTCAAGCGGGTGCATTACT       TTTGGTAAACCGGTTGTCTGGCTTGGCCATGAGGAAGCATCGCTGAAATCTCTCACA       TATTTTAATAGACCTAGTGTCGTTTGTGAAAATAAATTTAATGTGTTGCCCGTTGATG       TCAGTGAACCCACGGACAAGGGGCCTGTGCCTGCTGCAGTCCTTGTTACCGGCGTCC       CTGGAGCTGATGCGTCAGCTGGTGCCGGTATTGCCAAGGAGCAAAAAGCCTGTGCTT       CTGCTAGTGTGGAGGATCAGGTTGTTACGGAGGTTCGTCAAGAGCCATCTGTTTCAG       CTGCTGATGTCAAAGAGGTTAAATTGAATGGTGTTAAAAAGCCTGTTAAGGTGGAA       GGTAGTGTGGTTGTTAATGATCCCACTAGCGAAACCAAAGTTGTTAAAAGTTTGTCT       ATTGTTGATGTCTATGATATGTTCCTGACAGGGTGTAAGTATGTGGTTTGGACTGCTA       ATGAGTTGTCTCGACTAGTAAATTCACCGACTGTTAGGGAGTATGTGAAGTGGGGTA       AGGGAAAGATTGTAACACCCGCTAAGTTGTTGTTGTTAAGAGATGAGAAGCAAGAG       TTCGTAGCGCCAAAAGTAGTCAAGGCGAAAGCTATTGCCTGCTATTGTGCTGTGAAG       TGGTTTCTCCTCTATTGTTTTAGTTGGATAAAGTTTAATACTGATAATAAGGTTATAT       ACACCACAGAAGTAGCTTCAAAGCTTACTTTCAAGTTGTGCTGTTTGGCCTTTAAGA       ATGCCTTACAGACGTTTAATTGGAGCGTTGTGTCTAGGGGCTTTTTCCTAGTTGCAAC       GGTCTTTTTATTATGGTTTAACTTTTTGTATGCTAATGTTATTTTGAGTGACTTCTATT       TGCCTAATATTGGGCCTCTCCCTACGTTTGTGGGACAGATAGTTGCGTGGTTTAAGA       CTACATTTGGTGTGTCAACCATCTGTGATTTCTACCAGGTGACGGATTTGGGCTATA       GAAGTTCGTTTTGTAATGGAAGTATGGTATGTGAACTATGCTTCTCAGGTTTTGATAT       GCTGGACAACTATGATGCTATAAATGTTGTTCAACACGTTGTAGATAGGCGTTTGTC       CTTTGACTATATTAGCCTATTTAAATTAGTAGTTGAGCTTGTAATCGGCTACTCTCTT       TATACTGTGTGCTTCTACCCACTGTTTGTCCTTATTGGAATGCAGTTGTTGACCACAT       GGTTGCCTGAATTCTTTATGCTGGAGACTATGCATTGGAGTGCTCGTTTGTTTGTGTT       TGTTGCCAATATGCTTCCAGCTTTTACGTTACTGCGATTTTACATCGTGGTGACAGCT       ATGTATAAGGTCTATTGTCTTTGTAGACATGTTATGTATGGATGTAGTAAGCCTGGTT       GCTTGTTTTGTTATAAGAGAAACCGTAGTGTCCGTGTTAAGTGTAGCACCGTTGTTG       GTGGTTCACTACGCTATTACGATGTAATGGCTAACGGCGGCACAGGTTTCTGTACAA       AGCACCAGTGGAACTGTCTTAATTGCAATTCCTGGAAACCAGGCAATACATTCATAA       CTCATGAAGCAGCGGCGGACCTCTCTAAGGAGTTGAAACGCCCTGTGAATCCAACA       GATTCTGCTTATTACTCGGTCACAGAGGTTAAGCAGGTTGGTTGTTCCATGCGTTTGT       TCTACGAGAGAGATGGACAGCGTGTTTATGATGATGTTAATGCTAGTTTGTTTGTGG       ACATGAATGGTCTGCTGCATTCTAAAGTTAAAGGTGTGCCTGAAACGCATGTTGTGG       TTGTTGAGAATGAAGCTGATAAAGCTGGTTTTCTCGGCGCCGCAGTGTTTTATGCAC       AATCGCTCTACAGACCTATGTTGATGGTGGAAAAGAAATTAATAACTACCGCCAAC       ACTGGTTTGTCTGTTAGTCGAACTATGTTTGACCTTTATGTAGATTCATTGCTGAACG       TCCTCGACGTGGATCGCAAGAGTCTAACAAGTTTTGTAAATGCTGCGCACAACTCTC       TAAAGGAGGGTGTTCAGCTTGAACAAGTTATGGATACCTTTATTGGCTGTGCCCGAC       GTAAGTGTGCTATAGATTCTGATGTTGAAACCAAGTCTATTACCAAGTCCGTCATGT       CGGCAGTAAATGCTGGCGTTGATTTTACGGATGAGAGTTGTAATAACTTGGTGCCTA       CCTATGTTAAAAGTGACACTATCGTTGCAGCCGATTTGGGTGTTCTTATTCAGAATA       ATGCTAAGCATGTACAGGCTAATGTTGCTAAAGCCGCTAATGTGGCTTGCATTTGGT       CTGTGGATGCTTTTAACCAGCTATCTGCTGACTTACAGCATAGGCTGCGAAAAGCAT       GTTCAAAAACTGGCTTGAAGATTAAGCTTACTTATAATAAGCAGGAGGCAAATGTTC       CTATTTTAACTACACCGTTCTCTCTTAAAGGGGGCGCTGTTTTTAGTAGAATGTTACA       ATGGTTGTTTGTTGCTAATTTGATTTGTTTCATTGTGTTGTGGGCCCTTATGCCAACA       TATGCAGTGCACAAATCGGATATGCAGTTGCCTTTATATGCCAGTTTTAAAGTTATA       GATAATGGTGTGCTAAGGGATGTGTCTGTTACTGACGCATGCTTCGCAAACAAATTT       AATCAATTTGATCAATGGTATGAGTCTACTTTTGGTCTTGCTTATTACCGCAACTCTA       AGGCTTGTCCTGTTGTGGTTGCTGTAATAGATCAAGACATTGGCCATACCTTATTTAA       TGTTCCTACCACAGTTTTAAGATATGGATTTCATGTGTTGCATTTTATAACCCATGCA       TTTGCTACTGATAGCGTGCAGTGTTACACGCCACATATGCAAATCCCCTATGATAAT       TTCTATGCTAGTGGTTGCGTGTTGTCATCCCTCTGTACTATGCTTGCGCATGCAGATG       GAACCCCGCATCCTTATTGTTATACAGGGGGTGTTATGCACAATGCCTCTCTGTATA       GTTCTTTGGCTCCTCATGTCCGTTATAACCTGGCTAGTTCAAATGGTTATATACGTTT       TCCCGAAGTGGTTAGTGAAGGCATTGTGCGTGTTGTGCGCACTCGCTCTATGACCTA       CTGCAGGGTTGGTTTATGTGAGGAGGCCGAGGAGGGTATCTGCTTTAATTTTAATCG       TTCATGGGTATTGAACAACCCGTATTATAGGGCCATGCCTGGAACTTTTTGTGGTAG       GAATGCTTTTGATTTAATACATCAAGTTTTAGGAGGATTAGTGCGGCCTATTGATTTC       TTTGCCTTAACGGCGAGTTCAGTGGCTGGTGCTATCCTTGCAATTATTGTCGTTTTGG       CTTTCTATTATTTAATAAAGCTTAAACGTGCCTTTGGTGACTACACTAGTGTTGTGGT       TATCAATGTAATTGTGTGGTGTATAAATTTTCTGATCGTTTTTGTGTTTCAGGTTTATC       CCACATTGTCTTGTTTATATGCTTGTTTTTATTTCTACACAACGCTTTATTTCCCTTCG       GAGATAAGTGTTGTTATGCATTTGCAATGGCTTGTCATGTATGGTGCTATTATGCCCT       TGTGGTTTTGCATTATTTACGTGGCAGTCGTTGTTTCAAACCATGCATTGTGGTTGTT       CTCTTACTGCCGCAAAATTGGTACCGAGGTTCGTAGTGACGGCACATTTGAGGAAAT       GGCCCTTACTACCTTTATGATTACTAAAGAATCTTATTGTAAGTTGAAAAATTCTGTT       TCTGATGTTGCTTTTAACAGGTACTTGAGTCTTTATAACAAGTATCGTTATTTTAGTG       GCAAAATGGATACTGCCGCTTATAGAGAGGCTGCCTGTTCACAACTGGCAAAGGCA       ATGGAAACATTTAACCATAATAATGGTAATGATGTTCTCTATCAGCCTCCAACCGCC       TCTGTTACTACATCATTTTTACAGTCTGGTATAGTGAAGATGGTGTCGCCCACCTCTA       AAGTGGAGCCTTGTATTGTTAGTGTTACTTATGGTAACATGACACTTAATGGGTTGT       GGTTGGATGATAAAGTTTATTGCCCAAGACATGTTATCTGTTCTTCAGCTGACATGA       CAGACCCTGATTATCCTAATTTGCTTTGTAGAGTGACATCAAGTGATTTTTGTGTTAT       GTCTGGTCGTATGAGCCTTACTGTAATGTCTTATCAAATGCAGGGCTGCCAACTTGTT       TTGACTGTTACACTGCAAAATCCTAACACGCCTAAGTATTCCTTCGGTGTTGTTAAGC       CTGGTGAGACATTTACTGTACTGGCTGCATACAATGGCAGACCTCAAGGAGCCTTCC       ATGTTACGCTTCGTAGTAGCCATACCATAAAGGGCTCCTTTCTATGTGGATCCTGCG       GTTCTGTAGGATATGTTTTAACTGGCGATAGTGTACGATTTGTTTATATGCATCAGCT       AGAGTTGAGTACTGGTTGTCATACCGGTACTGACTTTAGTGGGAACTTTATATGGTCC       CTATAGAGATGCGCAAGTTGTACAATTGCCTGTTCAGGATTATACGCAGACTGTTAA       TGTTGTAGCTTGGCTTTATGCTGCTATTTTTAACAGATGCAACTGGTTTGTGCAAAGT       GATAGTTGTTCCCTGGAGGAGTTTAATGTTTGGGCTATGACCAATGGTTTTAGCTCA       ATCAAAGCCGATCTTGTCTTGGATGCGCTTGCTTCTATGACAGGCGTTACAGTTGAA       CAGGTGTTGGCCGCTATTAAGAGGCTGCATTCTGGATTCCAGGGCAAACAAATTTTA       GGTAGTTGTGTGCTTGAAGATGAGCTGACACCAAGTGATGTTTATCAACAACTAGCT       GGTGTCAAGCTACAGTCAAAGCGCACAAGAGTTATAAAAGGTACATGTTGCTGGAT       ATTGGCTTCAACGTTTTTGTTCTGTAGCATTATCTCAGCATTTGTAAAATGGACTATG       TTTATGTATGTTACTACCCATATGTTGGGAGTGACATTGTGTGCACTTTGTTTTGTAA       GCTTTGCTATGTTGTTGATCAAGCATAAGCATTTGTATTTAACTATGTATATTATGCC       TGTGTTATGCACACTGTTTTACACCAACTATTTGGTTGTGTACAAACAGAGTTTTAGA       GGTCTAGCTTATGCTTGGCTTTCACACTTTGTCCCTGCTGTAGATTATACATATATGG       ATGAAGTTTTATATGGTGTTGTGTTGCTAGTAGCTATGGTGTTTGTTACCATGCGTAG       CATAAACCACGACGTCTTTTCTATTATGTTTCTTGGTTGGTAGACTTGTCAGCCTGGTA       TCCATGTGGTATTTTGGAGCCAATTTAGAGGAAGAGGTACTATTGTTCCTCACATCC       CTATTTGGCACGTACACATGGACTACTATGTTGTCATTGGCTACCGCTAAGGTTATTG       CTAAATGGTTGGCTGTGAATGTCTTGTACTTCACAGACGTACCGCAAATTAAATTAG       TTCTTTTGAGCTACTTGTGTATTGGTTATGTGTGTTGTTGTTATTGGGGAATCTTGTCA       CTCCTTAATAGCATTTTTAGGATGCCATTGGGCGTCTACAATTATAAAATCTCCGTTC       AGGAGTTACGTTATATGAATGCTAATGGCTTGCGCCCACCTAGAAATAGTTTTGAGG       CCCTGATGCTTAATTTTAAGCTGTTGGGAATTGGTGGTGTGCCAGTCATTGAAGTAT       CTCAAATTCAATCAAGATTGACGGATGTTAAATGTGCTAATGTTGTGTTGCTTAATT       GCCTCCAGCACTTGCATATTGCATCTAATTCTAAGTTGTGGCAGTATTGTAGTACTTT       GCACAATGAAATACTGGCTACATCTGATTTGAGCGTGGCCTTCGATAAGTTGGCTCA       GCTCTTAGTTGTTTTATTTGCTAATCCAGCAGCAGTGGATAGCAAGTGCCTTGCAAG       TATTGAAGAAGTGAGCGATGATTACGTTCGCGACAATACTGTCTTGCAAGCCTTACA       GAGTGAATTTGTTAATATGGCTAGCTTCGTTGAGTATGAACTTGCTAAGAAGAATCT       AGATGAGGCTAAGGCTAGCGGCTCTGCCAATCAACAGCAGATTAAGCAGCTAGAGA       AGGCGTGTAATATTGCTAAGTCAGCATATGAGCGCGACAGAGCTGTTGCTCGTAAGC       TGGAACGTATGGCTGATTTAGCTCTTACAAACATGTATAAAGAAGCTAGAATTAATG       ATAAGAAGAGTAAGGTAGTGTCTGCATTGCAAACCATGCTCTTTAGTATGGTGCGTA       AGCTAGATAACCAAGCTCTTAATTCTATTTTAGATAATGCAGTTAAGGGTTGTGTAC       CTTTGAATGCAATACCATCATTGACTTCGAACACTCTGACTATAATAGTGCCAGATA       AGCAGGTTTTTGATCAGGTTGTGGATAATGTGTATGTCACCTATGCTGGGAATGTAT       GGCATATACAGTTTATTCAAGATGCTGATGGTGCTGTTAAACAATTGAATGAGATAG       ATGTTAATTCAACCTGGCCTCTAGTCATTGCTGCAAATAGGCATAATGAAGTGTCTA       CTGTTGTTTTGCAGAACAATGAGTTGATGCCTCAGAAGTTGAGAACTCAGGTTGTCA       ATAGTGGCTCAGATATGAATTGTAATACTCCTACCCAGTGTTACTATAATACTACTG       GCACGGGTAAGATTGTGTATGCTATACTTAGTGACTGTGATGGTCTCAAGTACACTA       AGATAGTAAAAGAAGATGGAAATTGTGTTGTTTTGGAATTGGATCCTCCCTGTAAGT       TTTCTGTTCAGGATGTGAAGGGCCTTAAAATTAAGTACCTTTACTTTGTGAAGGGGT       GTAATACACTGGCTAGAGGCTGGGTTGTAGGCACCTTATCCTCGACAGTGAGATTGC       AGGCGGGTACGGCAACTGAGTATGCCTCCAACTCTGCAATACTGTCGCTGTGTGCGT       TTTCTGTAGATCCTAAGAAAACGTACTTGGATTATATAAAACAGGGTGGAGTTCCCG       TTACTAATTGTGTTAAGATGTTATGTGACCATGCTGGCACTGGTATGGCCATTACTAT       TAAGCCGGAGGCAACCACTAATCAGGATTCTTATGGTGGTGCTTTCCGTTTGTATATA       TTGCCGCTCGCGTGTTGAACATCCAGATGTTGATGGATTGTGCAAATTACGCGGCAA       GTTTGTCCAAGTGCCCTTAGGCATAAAAGATCCTGTGTCATATGTGTTGACGCATGA       TGTTTGTCAGGTTTGTGGCTTTTGGCGAGATGGTAGCTGTTCCTGTGTAGGCACAGG       CTCCCAGTTTCAGTCAAAAGACACGAACTTTTTAAACGGGTTCGGGGTACAAGTGTA       AATGCCCGTCTTGTACCCTGTGCCAGTGGCTTGGACACTGATGTTCAATTAAGGGCA       TTTGACATTTGTAATGCTAATCGAGCTGGCATTGGTTTGTATTATAAAGTGAATTGCT       GCCGCTTCCAGCGTGTAGATGAGGACGGCAACAAGTTGGATAAGTTCTTTGTTGTTA       AAAGAACTAATTTAGAAGTGTATAATAAGGAGAAAGAATGCTATGAGTTGACAAAA       GAATGCGGTGTTGTGGCTGAACACGAGTTCTTCACATTTGATGTGGAGGGAAGTCGG       GTACCACACATAGTCCGTAAAGATCTTTCAAAGTTTACTATGTTAGATCTTTGCTATG       CATTGCGTCATTTTGACCGCAATGATTGTTCAACTCTTAAGGAAATTCTCCTTACATA       TGCTGAGTGTGAAGAGTCCTACTTCCAAAAGAAGGACTGGTATGATTTTGTTGAGAA       TCCTGATATAATTAATGTGTATAAAAAGCTTGGTCCTATATTTAATAGAGCCCTGCTT       AACACTGCCAAGTTTGCAGACGCATTAGTGGAGGCAGGCTTAGTAGGTGTTTTAACA       CTTGATAATCAAGATTTATATGGTCAATGGTATGACTTTGGAGATTTTGTCAAGACA       GTACCTGGTTGTGGTGTTGCCGTGGCAGACTCTTATTATTCATATATGATGCCAATGC       TGACTATGTGTCATGCGTTGGATAGTGAGTTGTTTGTTAATGGTACTTATAGGGAGTT       TGACCTTGTTCAGTATGATTTTACTGATTTCAAGCTAGAGCTCTTCACTAAGTATTTT       AAGCATTGGAGTATGACCTACCACCCGAACACCTGTGAGTGCGAGGATGACAGGTG       CATTATTCATTGCGCCAATTTTAATATACTTTTTAGTATGGTCTTACCTAAGACCTGT       TTTGGGCCTCTTGTTAGGCAGATATTTGTGGATGGTGTTCCTTTCGTTGTGTCGATCG       GTTACCATTATAAAGAATTAGGTGTTGTTATGAATATGGATGTGGATACACATCGTT       ATCGCTTGTCTCTTAAGGACTTGCTTTTGTATGCTGCAGACCCTGCCCTTCATGTGGC       GTCTGCTAGTGCACTGCTTGATTTGCGCACATGTTGTTTTAGCGTTGCAGCTATTACA       AGTGGCGTAAAATTTCAAACAGTTAAACCTGGAAATTTTAATCAGGATTTTTATGAG       TTTATTTTGAGTAAAGGCCTGCTTAAAGAGGGGAGCTCCGTTGATTTGAAGCACTTC       TTCTTTACGCAGGATGGTAATGCTGCTATTACTGATTATAATTATTACAAGTATAATC       TACCCACCATGGTGGATATTAAGCAGTTGTTGTTTGTTTTAGAAGTTGTTAATAAGTA       TTTTGAGATCTATGAGGGTGGGTGTATACCCGCAACACAGGTCATTGTTAATAATTA       TGATAAGAGTGCTGGCTATCCATTTAATAAATTTGGAAAGGCCAGGCTCTATTATGA       GGCATTATCATTTGAGGAGCAGGATGAAATTTATGCGTATACCAAACGCAATGTCCT       GCCGACCCTAACTCAAATGAATCTTAAATATGCTATTAGTGCTAAGAATAGGGCCCG       CACCGTTGCTGGTGTCTCTATTCTCAGTACTATGACTGGCAGAATGTTTCATCAAAA       GTGTCTAAAGAGTATAGCAGCTACTCGCGGTGTTCCTGTAGTTATAGGCACCACGAA       GTTCTATGGCGGTTGGGATGATATGTTACGCCGCCTTATTAAAGATGTTGATAGTCC       TGTACTCATGGGTTGGGACTATCCTAAATGTGATCGTGCTATGCCAAACATACTGCG       TATTGTTAGTAGTTTGGTGCTAGCCCGTAAACATGATTCGTGCTGTTCGCATACGGAT       AGATTCTATCGTCTTGCGAACGAGTGCGCCCAAGTTTTGAGTGAAATTGTTATGTGT       GGTGGTTGTTATTATGTTAAACCAGGTGGCACTAGTAGTGGGGATGCAACCACTGCT       TTTGCTAATTCTGTGTTTAACATTTGTCAAGCTGTTTCCGCCAATGTATGCTCGCTTA       TGGCATGCAATGGACACAAAATTGAAGATTTGAGTATACGCGAGTTACAAAAGCGC       CTATACTCTAATGTCTATCGTGCGGACCATGTTGACCCCGCATTTGTTAGTGAGTATT       ATGAGTTTTTAAATAAGCATTTTAGTATGATGATTTTGAGTGATGATGGTGTTGTGTG       TTATAATTCAGAGTTTGCGTCCAAGGGTTATATTGCTAATATAAGTGCCTTTCAACA       GGTATTATATTATCAAAATAATGTGTTTATGTCTGAGGCCAAATGTTGGGTAGAAAC       AGACATCGAAAAGGGACCGCATGAATTTTGTTCTCAACATACAATGCTAGTCAAGAT       GGATGGTGATGAAGTCTACCTTCCATACCCTGATCCTTCGAGAATCTTAGGAGCAGG       CTGTTTTGTTGATGATTTATTAAAGACTGATAGCGTTCTCTTGATAGAGCGTTTCGTA       AGTCTTGCAATTGATGCTTATCCTTTAGTATACCATGAGAACCCAGAGTATCAAAAT       GTGTTCCGGGTATATTTAGAATATATAAAGAAGCTGTACAATGATCTCGGTAATCAG       ATCCTGGACAGCTACAGTGTTATTTTAAGTACTTGTGATGGTCAAAAGTTTACTGAT       GAGACCTTTTACAAGAACATGTATTTAAGAAGTGCAGTGCTGCAAAGCGTTGGTGCC       TGCGTTGTCTGTAGTTCTCAAACATCATTACGTTGTGGCAGTTGCATACGCAAGCCTT       TGCTGTGTTGCAAATGCGCCTATGATCATGTTATGTCCACTGATCATAAATATGTCCT       GAGTGTGTCACCATATGTGTGTAATTCACCGGGATGTGATGTAAATGATGTTACCAA       ATTGTATTTAGGTGGTATGTCATATTATTGTGAGGACCATAAACCACAGTATTCATTC       AAATTGGTGATGAATGGTATGGTTTTTGGTTTATATAAACAATCTTGTACTGGTTCGC       CCTACATAGAGGATTTTAATAAAATAGCTAGTTGCAAATGGACAGAAGTCGATGATT       ATGTGCTAGCTAATGAATGCACCGAACGCCTTAAATTGTTTGCCGCAGAAACGCAGA       AGGCCACAGAAGAGGCCTTTAAGCAATGTTATGCGTCAGCAACGATCCGTGAGATC       GTGAGCGATCGGGAGTTAATTTTATCTTGGGAAATTGGTAAAGTGAGACCACCACTT       AATAAAAATTATGTTTTTACTGGCTACCATTTTACTAATAATGGTAAGACAGTTTTAG       GTGAGTATGTTTTTGATAAGAGTGAGTTGACTAATGGTGTGTACTATCGCGCCACAA       CCACTTATAAGTTATCTGTAGGTGATGTGTTCATTTTAACATCACACGCAGTGTCTAG       TTTAAGTGCTCCTACATTAGTACCGCAGGAGAATTATACTAGCATTCGTTTTGCTAGT       GTTTATAGTGTGCCTGAGACGTTTCAGAATAATGTGCCTAATTATCAGCACATTGGA       ATGAAGCGCTATTGTACTGTACAGGGACCGCCTGGTACTGGTAAGTCCCATCTAGCC       ATTGGGCTAGCTGTTTATTATTGTACAGCGCGCGTGGTGTATACCGCTGCTAGCCAT       GCTGCAGTTGACGCGCTGTGTGAAAAGGCACATAAATTTTTAAATATTAATGACTGC       ACGCGTATTGTTCCTGCAAAGGTGCGTGTAGATTGTTATGATAAATTTAAGGTCAAT       GACACCACTCGCAAGTATGTGTTTACTACAATAAATGCATTACCTGAGTTGGTGACT       GACATTATTGTCGTTGATGAAGTTAGTATGCTTACCAACTATGAGCTGTCTGTTATTA       ACAGTCGTGTTAGTGCTAAGCATTATGTGTATATTGGAGACCCTGCGCAGTTACCTG       CACCACGTGTGCTACTGAATAAGGGAACTCTAGAACCTAGATATTTTAATTCCGTTA       CCAAGCTAATGTGTTGTTTGGGTCCAGATATTTTCTTGGGCACCTGTTATAGATGCCC       TAAGGAGATTGTGGATACGGTGTCAGCCTTGGTTTATAATAATAAGCTGAAGGCTAA       AAATGATAATAGCTCCATGTGCTTTAAGGTTTATTATAAGGGCCAGACTACACATGA       GAGTTCTAGTGCTGTTAATATGCAGCAAATACATTAATTAGTAAGTTTTTAAAGGC       AAACCCCAGTTGGAGTAACGCCGTATTTATTAGTCCTTATAATAGTCAGAACTATGT       TGCTAAGAGAGTCTTGGGATTACAAACCCAGACAGTAGACTCAGCGCAGGGTTCTG       AATATGATTTTGTTATTTATTCACAGACTGCGGAAACAGCGCATTCTGTCAATGTAA       ATAGATTCAATGTTGCTATTACACGTGCTAAGAAGGGTATTCTCTGTGTCATGAGTA       GTATGCAATTATTTGAGTCTCTTAATTTTACTACACTGACGTTGGATAAGATTAACAA       TCCACGATTACAGTGTACTACAAATTTGTTTAAGGATTGTAGCAGGAGCTATGTAGG       ATATCACCCAGCCCATGCACCATCCTTTTTGGCAGTTGATGACAAATATAAGGTAGG       CGGTGATTTAGCCGTTTGCCTTAATGTTGCTGATTCTGCTGTCACTTATTCGCGGCTT       ATATCACTCATGGGATTCAAGCTTGACTTGACCCTTGATGGTTATTGTAAGCTGTTTA       TAACTAGAGATGAAGCTATCAAACGTGTTAGAGCCTGGGTTGGCTTCGATGCAGAA       GGTGCCCATGCGATACGTGATAGCATTGGGACAAATTTCCCATTACAATTAGGCTTT       TCGACTGGAATTGATTTTGTTGTCGAAGCCACTGGAATGTTTGCTGAGAGAGATGGT       TATGTCTTTAAAAAGGCAGCCGCACGAGCTCCTCCTGGCGAACAATTTAAACACCTT       ATCCCACTTATGTCAAGAGGGCAGAAATGGGATGTGGTTCGAATTAGAATAGTACA       AATGTTGTCAGACCACCTAGCGGATTTGGCAGACAGTGTTGTACTTGTGACGTGGGC       TGCCAGCTTTGAGCTCACATGTTTGCGATATTTCGCTAAAGTTGGAAGAGAAGTTGT       GTGTAGTGTCTGCACCAAGCGTGCGACATGTTTAAATTCTAGAACTGGATACTATGG       ATGCTGGCGACATAGTTATTCCTGTGATTACCTGTACAACCCACTAATAGTTGACAT       TCAACAGTGGGGATATACAGGATCTTTAACTAGCAATCATGATCCTATTTGCAGCGT       GCATAAGGGTGCTCATGTTGCATCATCTGATGCTATCATGACCCGGTGTCTAGCTGT       TCATGATTGCTTTTGTAAGTCTGTTAATTGGAATTTAGAATACCCCATTATTTCAAAT       GAGGTCAGTGTTAATACCTCCTGCAGGTTATTGCAGCGCGTAATGTTTAGGGCTGCG       ATGCTATGCAATAGGTATGATGTGTGTTATGACATTGGCAACCCTAAAGGTCTTGCC       TGTGTCAAAGGATATGATTTTAAGTTTTATGATGCCTCCCCTGTTGTTAAGTCTGTTA       AACAGTTTGTTTATAAATACGAGGCACATAAAGATCAATTTTTAGATGGTTTGTGTA       TGTTTTGGAACTGCAATGTGGATAAGTATCCAGCGAATGCAGTTGTGTGTAGGTTTG       ACACGCGTGTGTTGAACAAATTAAATCTCCCTGGCTGTAATGGTGGCAGTTTGTATG       TTAACAAACATGCATTCCACACCAGTCCCTTTACCCGGGCTGCCTTCGAGAATTTGA       AGCCTATGCCTTTCTTTTATTATTCAGATACGCCCTGTGTGTATATGGAAGGCATGGA       ATCTAAGCAGGTCGATTATGTCCCATTGAGAAGCGCTACATGCATCACAAGATGCAA       TTTAGGTGGCGCTGTTTGTTTAAAACATGCTGAGGAGTATCGTGAGTACCTTGAGTC       TTACAATACGGCAACCACAGCGGGTTTTACTTTTTGGGTCTATAAGACTTTTGATTTT       TATAACCTTTGGAATACTTTTACTAGGCTCCAAAGTTTAGAAAATGTAGTGTATAAT       TTGGTCAATGCTGGACACTTTGATGGCCGGGCGGGTGAACTGCCTTGTGCTGTTATA       GGTGAGAAAGTCATTGCCAAGATTCAAAATGAGGATGTCGTGGTCTTTAAAAATAA       CACGCCATTCCCCACTAATGTGGCTGTCGAATTATTTGCTAAGCGCAGTATTCGGCC       CCACCCCGAGCTTAAGCTCTTTAGAAATTTGAATATTGACGTGTGCTGGAGTCACGT       CCTTTGGGATTATGCTAAGGATAGTGTGTTTTGCAGTTCGACGTATAAGGTCTGCAA       ATACACAGATTTACAGTGCATTGAAAGCTTGAATGTACTTTTTGATGGTCGTGATAA       TGGTGCTCTTGAAGCTTTTAAGAAGTGCCGGAATGGCGTCTACATTAACACGACAAA       AATTAAAAGTCTGTCGATGATTAAAGGCCCACAACGTGCCGATTTGAATGGCGTAGT       TGTGGAGAAAGTTGGAGATTCTGATGTGGAATTTTGGTTTGCTGTGCGTAAAGACGG       TGACGATGTTATCTTCAGCCGTACAGGGAGCCTTGAACCGAGCCATTACCGGAGCCC       ACAAGGTAATCCGGGTGGTAATCGCGTGGGTGATCTCAGCGGTAATGAAGCTCTAG       CGCGTGGCACTATCTTTACTCAAAGCAGATTATTATCTTCTTTCACACCTCGATCAGA       GATGGAGAAAGATTTTATGGATTTAGATGATGATGTGTTCATTGCAAAATATAGTTT       ACAGGACTACGCGTTTGAACACGTTGTTTATGGTAGTTTTAACCAGAAGATTATTGG       AGGTTTGCATTTGCTTATTGGCTTAGCCCGTAGGCAGCAAAAATCCAATCTGGTAAT       TCAAGAGTTCGTGACATACGACTCTAGCATTCATTCGTACTTTATCACTGACGAGAA       CAGTGGTAGTAGTAAGAGTGTGTGCACTGTTATTGATTTATTGTTAGATGATTTTGTG       GACATTGTAAAGTCCCTGAATCTAAAGTGTGTGAGTAAGGTTGTTAATGTTAATGTT       GATTTTAAAGATTTCCAGTTTATGTTGTGGTGCAATGAGGAGAAGGTCATGACTTTC       TATCCTCGTTTGCAGGCTGCTGCTGACTGGAAACCTGGTTATGTTATGCCTGTCTTAT       ATAAGTATTTGGAATCGCCTCTGGAAAGAGTAAACCTCTGGAATTATGGCAAGCCG       ATTACTTTACCTACAGGATGTATGATGAATGTTGCTAAGTATACTCAATTATGTCAAT       ATTTGAGCACTACAACATTAGCAGTTCCGGCTAATATGCGTGTCTTACACCTTGGTG       CCGGTTCGGATAAGGGTGTTGCCCCTGGGTCTGCAGTTCTTAGGCAGTGGCTACCAG       CGGGAAGTATTCTTGTAGATAATGATGTGAATCCATTTGTGAGTGACAGTGTCGCCT       CATATTATGGAAATTGTATAACCTTACCCTTTGATTGTCAGTGGGATCTGATAATTTC       TGATATGTACGACCCTCTTACTAAGAACATTGGGGAGTACAACGTGAGTAAAGATG       GATTCTTTACTTACCTCTGTCATTTAATTCGTGACAAGTTGGCTCTGGGTGGCAGTGT       TGCCATAAAAATAACAGAGTTTTCTTGGAACGCTGAGTTATATAGTTTAATGGGGAA       GTTTGCGTTCTGGACAATCTTTTGCACCAACGTAAACGCCTCTTCAAGTGAAGGAAA       TTTGATTGGCATAAATTGGTTGAATAAGACCCGTACCGAAATTGACGGTAAAACCAT       GCATGCCAATTATCTGTTTTGGAGAAATAGTACAATGTGGAATGGAGGGGCTTACAG       TCTCTTTGACATGAGTAAGTTCCCTTTGAAAGCGGCTGGTACGGCTGTTGTTAGCCTT       AAACCAGACCAAATAAATGACTTAGTCCTCTCCTTGATTGAGAAGGGCAAGTTATTA       GTGCGTGATACACGCAAAGAAGTTTTTGTTGGCGATAGCCTAGTAAATGTCAAATAA       ATCTATACTTGTCGTGGCTGTGAAAATGGCCTTTGCTGACAAGCCTAATCATTTCATA       AACTTTCCCCTGGCCCAATTTAGTGGCTTTATGGGTAAGTATTTAAAGCTACAGTCTC       AACTTGTGGAAATGGGTTTAGACTGTAAATTACAGAAGGCACCACATGTTAGTATTA       CCCTGCTTGATATTAAAGCAGACCAATACAAACAGGTGGAATTTGCAATACAAGAA       ATAATAGATGATCTGGCGGCATATGAGGGAGATATTGTCTTTGACAACCCTCACATG       CTTGGCAGATGCCTTGTTCTTGATGTTAGAGGATTTGAAGAGTTGCATGAAGATATT       GTTGAAATTCTCCGCAGAAGGGGTTGCACGGCAGATCAATCCAGACACTGGATTCC       GCACTGCACTGTGGCCCAATTTGACGAAGAAAGAGAAACAAAAGGAATGCAATTCT       ATCATAAAGAACCCTTCTACCTCAAGCATAACAACCTATTAACGGATGCTGGGCTTG       AGCTCGTGAAGATAGGTTCTTCCAAAATAGATGGGTTTTATTGTAGTGAACTGAGTG       TTTGGTGTGGTGAGAGGCTTTGTTATAAGCCTCCAACACCCAAATTCAGTGATATAT       TTGGCTATTGCTGCATAGATAAAATACGTGGTGATTTAGAAATAGGAGACCTACCGC       AGGATGATGAGGAAGCGTGGGCCGAGCTAAGTTACCACTATCAAAGAAACACCTAC       TTCTTCAGACATGTGCACGATAATAGCATCTATTTTCGTACCGTGTGTAGAATGAAG       GGTTGTATGTGTTGATTTGTTTTTACACTATTAGTGTAATAAGCTTATTATTTTGTTGA       AAAGGGCAGGATGTGCATAGCTATGGCTCCTCGCACACTGCTTTTGCTGATTTGATG       TCAGCTGGTGTTTGGGTTCAATGAACCTCTTAACATCGTTTCACATTTAAATGATGAC       TGGTTTCTATTTGGTGACAGTCGTTCTGACTGTACCTATGTAGAAAATAACGGTCATC       CTAAATTAGATTGGCTTGACCTCGACCCAAAGTTGTGTAATTCAGGAAAGATTTCCG       CAAAGAGTGGTAACTCTCTCTTTAGGAGTTTTCACTTCACTGATTTTTACAATTATAC       GGGTGAGGGAGACCAAATTGTATTTTATGAAGGAGTTAATTTTAGTCCCAGCCATGG       CTTTAAATGCCTGGCTCATGGAGATAATAAAAGATGGATGGGCAATAAAGCTCGAT       TTTATGCCCGAGTGTATGAGAAGATGGCCCAATATAGGAGCCTATCGTTTGTTAATG       TGTCTTATGCCTATGGAGGTAATGCAAAGCCCGCCTCCATTTGCAAAGACAATACTT       TAACACTCAATAACCCCACCTTCATATCGAAGGAGTCTAATTATGTTGATTATTACT       ATGAGAGTGAGGCTAATTTCACACTAGAAGGTTGTGATGAATTTATAGTACCGCTCT       GTGGTTTTAATGGCCATTCCAAGGGCAGCTCTTCGGATGCTGCCAATAAATATTATA       CTGACTCTCAGAGTTACTATAATATGGATATTGGTGTCTTATATGGGTTCAATTCGAC       CTTGGATGTTGGCAACACTGCTAAGGATCCGGGTCTTGATCTCACTTGCAGGTATCT       TGCATTGACTCCTGGTAATTATAAGGCTGTGTCCTTAGAATATTTGTTAAGCTTACCC       TCAAAGGCTATTTGCCTCCATAAGACAAAGCGCTTTATGCCTGTGCAGGTAGTTGAC       TCAAGGTGGAGTAGCATCCGCCAGTCAGACAATATGACCGCTGCAGCCTGTCAGCT       GCCATATTGTTTCTTTCGCAACACATCTGCGAATTATAGTGGTGGCACACATGATGC       GCACCATGGTGATTTTCATTTCAGGCAGTTATTGTCTGGTTTGTTATATAATGTTTCC       TGTATTGCCCAGCAGGGTGCATTTCTTTATAATAATGTTAGTTCCTCTTGGCCAGCCT       ATGGGTACGGTCATTGTCCAACGGCAGCTAACATTGGTTATATGGCACCTGTTTGTA       TCTATGACCCTCTCCCGGTCATACTGCTAGGTGTGTTATTGGGTATAGCTGTGTTGAT       TATTGTGTTTTTGATGTTTTATTTTATGACGGATAGCGGTGTTAGATTGCATGAGGCA       TAATCTAAACATGCTGTTCGTGTTTATTCTATTTTTGCCCTCTTGTTTAGGGTATATTG       GTGATTTTAGATGTATCCAGCTTGTGAATTCAAACGGTGCTAATGTTAGTGCTCCAA       GCATTAGCACTGAGACCGTTGAAGTTTCACAAGGCCTGGGGACATATTATGTGTTAG       ATCGAGTTTATTTAAATGCCACATTATTGCTTACTGGTTACTACCCGGTCGATGGTTC       TAAGTTTAGAAACCTCGCTCTTACGGGAACTAACTCAGTTAGCTTGTCGTGGTTTCA       ACCACCCTATTTAAGTCAGTTTAATGATGGCATATTTGCGAAGGTGCAGAACCTTAA       GACAAGTACGCCATCAGGTGCAACTGCATATTTTCCTACTATAGTTATAGGTAGTTT       GTTTGGCTATACTTCCTATACCGTTGTAATAGAGCCATATAATGGTGTTATAATGGCC       TCAGTGTGCCAGTATACCATTTGTCTGTTACCTTACACTGATTGTAAGCCTAACACTA       ATGGTAATAAGCTTATAGGGTTTTGGCACACGGATGTAAAACCCCCAATTTGTGTGT       TAAAGCGAAATTTCACGCTTAATGTTAATGCTGATGCATTTTATTTTCATTTTTACCA       ACATGGTGGTACTTTTTATGCGTACTATGCGGATAAACCCTCCGCTACTACGTTTTTG       TTTAGTGTATATATTGGCGATATTTTAACACAGTATTATGTGTTACCTTTCATCTGCA       ACCCAACAGCTGGTAGCACTTTTGCTCCGCGCTATTGGGTTACACCTTTGGTTAAGC       GCCAATATTTGTTTAATTTCAACCAGAAGGGTGTCATTACTAGTGCTGTTGATTGTGC       TAGTAGTTATACCAGTGAAATAAAATGTAAGACCCAGAGCATGTTACCTAGCACTG       GTGTCTATGAGTTATCCGGTTATACGGTCCAACCAGTTGGAGTTGTATACCGGCGTG       TTGCTAACCTCCCAGCTTGTAATATAGAGGAGTGGCTTACTGCTAGGTCAGTCCCCT       CCCCTCTCAACTGGGAGCGTAAGACTTTTCAGAATTGTAATTTTAATTTAAGCAGCC       TGTTACGTTATGTTCAGGCTGAGAGTTTGTTTTGTAATAATATCGATGCTTCCAAAGT       GTATGGCAGGTGCTTTGGTAGTATTTCAGTTGATAAGTTTGCTGTACCCCGAAGTAG       GCAAGTTGATTTACAGCTTGGTAACTCTGGATTTCTGCAGACTGCTAATTATAAGAT       TGATACAGCTGCCACTTCGTGTCAGCTGCATTACACCTTGCCTAAGAATAATGTCAC       CATAAACAACCATAACCCCTCGTCTTGGAATAGGAGGTATGGCTTTAATGATGCTGG       CGTCTTTGGCAAAAACCAACATGACGTTGTTTACGCTCAGCAATGTTTTACTGTAAG       ATCTAGTTATTGCCCGTGTGCTCAACCGGACATAGTTAGCCCTTGCACTACTCAGAC       TAAGCCTAAGTCTGCTTTTGTTAATGTGGGTGACCATTGTGAAGGCTTAGGTGTTTTA       GAAGATAATTGTGGCAATGCTGATCCACATAAGGGTTGTATCTGTGCCAACAATTCA       TTTATTGGATGGTCACATGATACCTGCCTTGTTAATGATCGCTGCCAAATTTTTGCTA       ATATATTGTTAAATGGCATTAATAGTGGTACCACATGTTCCACAGATTTGCAGTTGC       CTAATACTGAAGTGGTTACTGGCATTTGTGTCAAATATGACCTCTACGGTATTACTG       GACAAGGTGTTTTTAAAGAGGTTAAGGCTGACTATTATAATAGCTGGCAAACCCTTC       TGTATGATGTTAATGGTAATTTGAATGGTTTTCGTGATCTTACCACTAACAAGACTTA       TACGATAAGGAGCTGTTATAGTGGCCGTGTTTCTGCTGCATTTCATAAAGATGCACC       CGAACCGGCTCTGCTCTATCGTAATATAAATTGTAGCTATGTTTTTAGCAATAATATT       TCCCGTGAGGAGAACCCACTTAATTACTTTGATAGTTATTTGGGTTGTGTTGTTAATG       CTGATAACCGCACGGATGAGGCGCTTCCTAATTGTGATCTCCGTATGGGTGCTGGCT       TATGCGTTGATTATTCAAAATCACGCAGGGCTGACCGATCAGTTTCTACTGGCTATC       GGTTAACTACATTTGAGCCATACACTCCGATGTTAGTTAATGATAGTGTCCAATCCG       TTGATGGATTATATGAGATGCAAATACCAACCAATTTTACTATTGGGCACCATGAGG       AGTTCATTCAAACTAGATCTCCAAAGGTGACTATAGATTGTGCTGCATTTGTCTGTG       GTGATAACACTGCATGCAGGCAGCAGTTGGTTGAGTATGGCTCTTTCTGTGTTAATG       TTAATGCCATTCTTAATGAGGTTAATAACCTCTTGGATAATATGCAACTACAAGTTG       CTAGTGCATTAATGCAGGGTGTTACTATAAGCTCGAGACTGCCAGACGGCATCTCAG       GCCCTATAGATGACATTAATTTTAGTCCTCTACTTGGATGCATAGGTTCAACATGTGC       TGAAGACGGCAATGGACCTAGTGCAATCCGAGGGCGTTCTGCTATAGAGGATTTGTT       ATTTGACAAGGTCAAATTATCTGATGTTGGCTTTGTCGAGGCTTATAATAATTGCAC       CGGTGGTCAAGAAGTTCGTGACCTCCTTTGTGTACAATCTTTTAATGGCATCAAAGT       ATTACCTCCTGTGTTGTCAGAGAGTCAGATCTCTGGCTACACAACCGGTGCTACTGC       GGCAGCTATGTTCCCACCGTGGTCAGCAGCTGCCGGTGTGCCATTTAGTTTAAGTGT       TCAATATAGAATTAATGGTTTAGGTGTCACTATGAATGTGCTTAGTGAGAACCAAAA       GATGATTGCTAGTGCTTTTAACAATGCGCTGGGTGCTATCCAGGATGGGTTTGATGC       AACCAATTCTGCTTTAGGTAAGATCCAGTCCGTTGTTAATGCAAATGCTGAAGCACT       CAATAACTTACTAAATCAACTTTCTAACAGGTTTGGTGCTATTAGTGCTTCTTTACAA       GAAATTCTAACTCGGCTTGAGGCTGTAGAAGCAAAAGCCCAGATAGATCGTCTTATT       AATGGCAGGTTAACTGCACTTAATGCGTATATATCCAAGCAACTTAGTGATAGTACG       CTTATTAAAGTTAGTGCTGCTCAGGCCATAGAAAAGGTCAATGAGTGCGTTAAGAGC       CAAACCACGCGTATTAATTTCTGTGGCAATGGTAATCATATATTATCTCTTGTCCAGA       ATGCGCCTTATGGCTTATATTTTATACACTTCAGCTATGTGCCAATATCCTTTACAAC       CGCAAATGTGAGTCCTGGACTTTGCATTTCTGGTGATAGAGGATTAGCACCTAAAGC       TGGATATTTTGTTCAAGATGATGGAGAATGGAAGTTCACAGGCAGTTCATATTACTA       CCCTGAACCCATTACAGATAAAAACAGTGTCATTATGAGTAGTTGCGCAGTAAACTA       CACAAAGGCACCTGAAGTTTTCTTGAACACTTCAATACCTAATCCACCCGACTTTAA       GGAGGAGTTAGATAAATGGTTTAAGAATCAGACGTCTATTGCGCCTGATTTATCTCT       CGATTTCGAGAAGTTAAATGTTACTTTGCTGGACCTGACGTATGAGATGAACAGGAT       TCAGGATGCAATTAAGAAGTTAAATGAGAGCTACATCAACCTCAAGGAAGTTGGCA       CATATGAAATGTATGTGAAATGGCCTTGGTATGTTTGGTTGCTAATTGGATTAGCTG       GTGTAGCTGTTTGTGTGTTGTTATTCTTTATATGTTGCTGCACAGGTTGTGGCTCATG       TTGTTTTAAGAAGTGTGGAAATTGTTGTGATGAGTATGGAGGACACCAGGACAGTAT       TGTGATACATAATATTTCCTCTCATGAGGATTGACTATCACAGCCTCTCCTGGAAAG       ACAGAAAATCTAAACAATTTATAGCATTCTCATTGCTACCTGGCCCCGTAAGAGGCA       GTCATAGCTATGGCCGTGTTGGTCCTAAGGCTACATTGGCTGCTGTCTTTATTGGTCC       ATTTATTGTAGCATGTATGCTAGGCATTGGCCTAGTTTATTTATTGCAATTGCAAGTT       CAAATTTTTCATGTTAAGGATACCATACGTGTGACTGGCAAGCCAGCCACTGTGTCT       TATACTACAAGTACACCAGTAACACCGAGCGCGACGACGCTCGATGGTACTACGTA       TACTTTAATTAGACCCACTAGCTCTTATACAAGAGTTTATCTTGGTACTCCAAGAGGT       TTTGATTATAGTACATTTGGGCCTAAGACCCTAGATTATGTTACTAATCTAAACCTCA       TCTTAATTCTGGTCGTCCATATACTTTTAAGGCATTGTCCAGGCATATGAGACCAAC       AGCCACATGGATTTGGCATGTGAGTGATGCATGGTTACGCCGCACGCGGGACTTTGG       TGTCATTCGCCTAGAAGATTTTTGTTTTCAATTTAATTATAGCCAACCCCGAGTTGGT       TATTGTAGAGTTCCTTTAAAGGCTTGGTGTAGCAACCAGGGTAAATTTGCAGCGCAG       TTTACCCTAAAAAGTTGCGAAAAACCAGGTCACGAAAAATTTATTACTAGCTTCACG       GCCTACGGCAGAACTGTCCAACAGGCCGTTAGCAAGTTAGTAGAAGAAGCTGTTGA       TTTTATTCTTTTTAGGGCCACGCAGCTCGAAAGAAATGTTTAATTTATTCCTTACAGA       CACAGTATGGTATGTGGGGCAGATTATTTTTATATTCGCAGTGTGTTTGATGGTCACC       ATAATTGTGGTTGCCTTCCTTGCGTCTATCAAACTTTGTATTCAACTTTGCGGTTTAT       GTAATACTTTGGTGCTGTCCCCTTCTATTTATTTGTATGATAGGAGTAAGCAGCTTTA       TAAGTATTATAATGAAGAAATGAGACTGCCCCTATTAGAGGTGGATGATATCTAATC       TAAACATTATGAGTAGTACTACTCAGGCCCCAGAGCCCGTCTATCAATGGACGGCCG       ACGAGGCAGTTCAATTCCTTAAGGAATGGAACTTCTCGTTGGGCATTATACTACTCT       TTATTACTATCATACTACAGTTCGGTTACACGAGCCGTAGCATGTTTATTTATGTTGT       GAAAATGATAATCTTGTGGTTAATGTGGCCACTGACTATTGTTTTGTGTATTTTCAAT       TGCGTGTATGCGCTAAATAATGTGTATCTTGGATTTTCTATAGTGTTTACTATAGTGT       CCATTGTAATCTGGATTATGTATTTTGTTAATAGCATAAGGTTGTTTATCAGGACTGG       TAGCTGGTGGAGCTTCAACCCCGAAACAAACAACCTTATGTGTATAGATATGAAAG       GTACCGTGTATGTTAGACCCATTATTGAGGATTACCATACACTAACAGCCACTATTA       TTCGTGGCCACCTCTACATGCAAGGTGTTAAGCTAGGCACCGGTTTCTCTTTGTCTGA       CTTGCCCGCTTATGTTACAGTTGCTAAGGTGTCACACCTTTGCACTTATAAGCGCGCA       TTCTTAGACAAGGTAGACGGTGTTAGCGGTTTTGCTGTTTATGTGAAGTCCAAGGTC       GGAAATTACCGACTGCCCTCAAACAAACCGAGTGGCGCGGACACCGCATTGTTGAG       AATCTAATCTAAACTTTAAGGATGTCTTTTGTTCCTGGGCAAGAAAATGCCGGTGGC       AGAAGCTCCTCTGTAAACCGCGCTGGTAATGGAATCCTCAAGAAGACCACTTGGGCT       GACCAAACCGAGCGTGGACCAAATAATCAAAATAGAGGCAGAAGGAATCAGCCAA       AGCAGACTGCAACTACTCAACCCAACTCCGGGAGTGTGGTTCCCCATTACTCCTGGT       TTTCTGGCATTACCCAGTTCCAAAAGGGAAAGGAGTTTCAGTTTGCAGAAGGACAA       GGAGTGCCTATTGCCAATGGAATCCCCGCTTCAGAGCAAAAGGGATATTGGTATAG       ACACAACCGCCGTTCTTTTAAAACACCTGATGGGCAGCAGAAGCAATTACTGCCCA       GATGGTATTTTTACTATCTTGGCACAGGGCCCCATGCTGGAGCCAGTTATGGAGACA       GCATTGAAGGTGTCTTCTGGGTTGCAAACAGCCAAGCGGACACCAATACCCGCTCTG       ATATTGTCGAAAGGGACCCAAGCAGTCATGAGGCTATTCCTACTAGGTTTGCGCCCG       GCACGGTATTGCCTCAGGGCTTTTATGTTGAAGGCTCTGGAAGGTCTGCACCTGCTA       GCCGATCTGGTTCGCGGTCACAATCCCGTGGGCCAAATAATCGCGCTAGAAGCAGTT       CCAACCAGCGCCAGCCTGCCTCTACTGTAAAACCTGATATGGCCGAAGAAATTGCTG       CTCTTGTTTTGGCTAAGCTCGGTAAAGATGCCGGCCAGCCCAAGCAAGTAACGAAGC       AAAGTGCCAAAGAAGTCAGGCAGAAAATTTTAAACAAGCCTCGCCAAAAGAGGACT       CCAAACAAGCAGTGCCCAGTGCAGCAGTGTTTTGGAAAGAGAGGCCCCAATCAGAA       TTTTGGAGGCTCTGAAATGTTAAAACTTGGAACTAGTGATCCACAGTTCCCCATTCTT       GCAGAGTTGGCTCCAACAGTTGGTGCCTTCTTCTTTGGATCTAAATTAGAATTGGTC       AAAAAGAATTCTGGTGGTGCTGATGAACCCACCAAAGATGTGTATGAGCTGCAATA       TTCAGGTGCAGTTAGATTTGATAGTACTCTACCTGGTTTTGAGACTATCATGAAAGT       GYGAATGAGAATTTGAATGCCTACCAGAAGGATGGTGGTGCAGATGTGGTGAGCC       CAAAGCCCCAAAGAAAAGGGCGTAGACAGGCTCAGGAAAAGAAAGATGAAGTAGA       TAATGTAAGCGTTGCAAAGCCCAAAAGCTCTGTGCAGCGAAATGTAAGTAGAGAAT       TAACCCCAGAGGATAGAAGTCTGTTGGCTCAGATCCTTGATGATGGCGTAGTGCCAG       ATGGGTTAGAAGATGACTCTAATGTGTAAAGAGAATGAATCCTATGTCGGCGCTCG       GTGGTAACCCCTCGCGAGAAAGTCGGGATAGGACACTCTCTATCAGAATGGATGTCT       TGCTGTCATAACAGATAGAGAAGGTTGTGGCAGACCCTGTATCAATTAGTTGAAAG       AGATTGCAAAATAGAGAATGTGTGAGAGAAGTTAGCAAGGTCCTACGTCTAACCAT       AAGAACGGCGATAGGCGCCCCCTGGGAAGAGCTCACATCAGGGTACTATTCCTGCA       ATGCCCTAGTAAATGAATGAAGTTGATCATGGCCAATTGGAAGAATCACAAAAAAA       AAAAAAAAAAAAAAAA          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 TGAACCCACCAAAGATGTGTATGAG 
                 (SEQ ID: No. 35) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CCATCCTTCTGGTAGGCATTCAAAT 
                 (SEQ ID: No. 36) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CTGCACCTGAATATTG 
                 (SEQ ID: No. 37) 
               
            
           
         
       
     
     
       
         
           
               
               
               
            
               
                   
               
               
                 GGCAGCTGCTGCTCCGAGGCGGTCAAGAGCGCCATGAGCACCATTGACCTGGACTC 
                 (SEQ ID: No. 38) 
                   
               
               
                 GCTGATGGCAGAGCACAGCGCTGCCTGGTACATGCCCGCTGACAAGGCCCTGGTGG 
               
               
                 ACAGCGCGGACGACGACAAGACGTTGGCGCCCTGGGAGAAGGCCAAACCCCAGAA 
               
               
                 CCCCAACAGCAAAGAAGGCTTGCAGCCAATTTACTGGAGCAGGGATGACGTAGCCC 
               
               
                 AGTGGCTCAAGTGGGCTGAAAATGAGTTTTCTTTAAGGCCAATTGACAGCAACACGT 
               
               
                 TTGAAATGAATGGCAAAGCTCTCCTGCTGCTGACCAAAGAGGACTTTCGCTATCGAT 
               
               
                 CTCCTCATTCAGGTGATGTGCTCTATGAACTCCTTCAGCATATTCTGAAGCAGAGGA 
               
               
                 AACCTCGGATTCTTTTTTCACC 
               
            
           
         
       
     
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 AAACCCCAGAACCCCAACAG 
                 (SEQ ID: No. 39) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 TCATCCCTGCTCCAGTAAATTGG 
                 (SEQ ID: No. 40) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CTGCAAGCCTTCTTTG 
                 (SEQ ID: No. 41) 
               
            
           
         
       
     
      mutation—a heritable change in DNA sequence resulting from mutagens. Various types of mutations including frame-shift mutations, missense mutations, and nonsense mutations.  
      Neomycin  
                          (SEQ ID: No. 42)                         CATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGA                   GGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCC               GCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGAC               CGACCTGTCCGGTGCCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTAT               CGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTC               ACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGA               TCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTG               ATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGAC               CACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGG               TCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAG               CCGAACTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTC               GTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGG               CCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCT               ATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGC               GAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTC               GCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTG          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 GGGCGCCCGGTTCTT 
                 (SEQ ID: No. 43) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CCTCGTCCTGCAGTTCATTCA 
                 (SEQ ID: No. 44) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 ACCTGTCCGGTGCCC 
                 (SEQ ID: No. 45) 
               
            
           
         
       
     
     
       
         
           
               
               
            
               
                   
               
               
                 (SEQ ID NO.: 46) 
                   
               
            
           
           
               
               
            
               
                 TTAAAGCTCATGCCTAGACCTGATGCTATAGAAGGTGTGCTCCTCGCTTC 
                   
               
               
                   
               
               
                 TCTGCCAATCTTAAGGTGCCCTGGATGGAGCTGGGTGACGTGTTTACCCT 
               
               
                   
               
               
                 TGTAGTCTGTCCTGTCTATATGCATGGATATGCACAGTGCCCTTGACCCA 
               
               
                   
               
               
                 ACCCTGCCAACCAGGCACCTGCAGAAGGTGTAGATGACCGTCAGATTGCC 
               
               
                   
               
               
                 CAGCATCCCTGTGAGTCCCACCAGCAGGATCACCGTGCCTAGGGTATAGT 
               
               
                   
               
               
                 GAGCATGGTCTGGGACATCGACTGTGGGGAAGGGGACCCAGGCAGCAGCC 
               
               
                   
               
               
                 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 
               
               
                   
               
               
                 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 
               
               
                   
               
               
                 NNNNNNNNNNNNNNNNNNNNNAGCCCATAGAAGAAAGTGCAAGTCTTCCA 
               
               
                   
               
               
                 AAATTTAACCCCACGCCCATATATGTGTGGATACTGAGCTTCTAAGAGGG 
               
               
                   
               
               
                 AGTGAAAGGCTCAGATGGCCTGCTGGAGGTTAACAGGACAAATGCGTGCC 
               
               
                   
               
               
                 TGCAGGACAGAGCACAGCTTGGGTGACCTTAAGGAATGAGTAGAGCCAGG 
               
               
                   
               
               
                 TCCTGGGTACTGCCCTCCCAACGAATGGATACCCCACAGCAAGCCTCCAA 
               
               
                   
               
               
                 GGAGAACTTGCAACCCCTGTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 
               
               
                   
               
               
                 NNAAACGAGGGAGGAGAACTTTCCACTAGAAAGAGAGTTTAGGTTCCCCC 
               
               
                   
               
               
                 AGGCTGCTGGGAGGCCATTTCCCCCATGAGGTTAGTACACAGGGACTAAG 
               
               
                   
               
               
                 GATAGCTCCCAGGGAGAGGCAGGAGTCTGCCCAATGTCCTGCCCAGCATC 
               
               
                   
               
               
                 CCACTCTGGCCTGTACAAGTCCAGAAGCCTAGGGCATGCCTTTCCCCCTA 
               
               
                   
               
               
                 GGATACTCCCCCAGGGGATNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 
               
               
                   
               
               
                 NNNNGAAGAGCAGGTCAGCCCCTGCCTTTCTGGTTCTCCAGTGGTCTCTG 
               
               
                   
               
               
                 CCAACAAAGACATTGCCTGTGCCCTCTTGTCTCAGCCACTGTGTAGAGAA 
               
               
                   
               
               
                 AGCTTAGAGAACTTCAGTGACGCTCAAGGTCCTTCGTCTAAGCTCAGACC 
               
               
                   
               
               
                 TTTTCTATCTCCCTGTTAAAACAAGGGTGGGGACAGGAGTCTCTGTGTAC 
               
               
                   
               
               
                 ACACATGCTCCCCAAACTTACCGTGGGGCTAACAGAGAGAAGCTGGGCTC 
               
               
                   
               
               
                 TTACGGAGACGTTCTGAGTGCCGTTCCAAATGCCTTGCAGGGCAGGACTG 
               
               
                   
               
               
                 GTTGTGAAGCTGGGATCCTGAGTTAAGCTTGACAAGAC 
               
            
           
         
       
     
     
       
         
           
               
               
               
            
               
                   
               
               
                 Forward Primer: 
                   
                   
               
               
                 TGGGTGACCTTAAGGAATGAGTAGA 
                 (SEQ ID: No. 47) 
               
               
                   
               
               
                 Reverse Primer: 
               
               
                 GTTCTCCTTGGAGGCTTGCT 
                 (SEQ ID: No. 48) 
               
               
                   
               
               
                 Probe: 
               
               
                 CTGCCCTCCCAACGAA 
                 (SEQ ID: No. 49) 
               
            
           
         
       
     
     
       
         
           
               
               
            
               
                   
               
               
                 (SEQ ID: No. 50) 
                   
               
            
           
           
               
               
            
               
                 GTGATGATGATGGGCAACGTTCACGTAGCAGCTCTTCTGCTCAACTACGG 
                   
               
               
                   
               
               
                 TGCAGATTCGAACTGCGAGGACCCCACTACCTTCTCCCGCCCGGTGCACG 
               
               
                   
               
               
                 ACGCAGCGCGGGAAGGCTTCCTGGACACGCTGGTGGTGCTGCACGGGTCA 
               
               
                   
               
               
                 GGGGCTCGGCTGGATGTGCGCGATGCCTGGGGTCGCCTGCCGCTCGACTT 
               
               
                   
               
               
                 GGCCCAAGAGCGGGGACATCAAGACATCGTGCGATATTTGCGTTCCGCTG 
               
               
                   
               
               
                 GGTGCTCTTTGTGTTCCGCTGGGTGGTCTTTGTGTACCGCTGGGAACGTC 
               
               
                   
               
               
                 GCCCAGACCGACGGGCATAGCTTCAGCTCAAGCACGCCCAG 
               
            
           
         
       
     
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 CGAGGACCCCACTACCTTCT 
                 (SEQ ID: No. 51) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CCGCTCTTGGGCCAAGT 
                 (SEQ ID: No. 52) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CAGGCATCGCGCACAT 
                 (SEQ ID: No. 53) 
               
            
           
         
       
     
      plate controls—are wells that include the house-keeping probe without nucleic acid sample.  
      Puromycin Sequence  
                          (SEQ ID: No. 54)                         ATGACCGAGTACAAGCCCACGGTGCGCCTCGCCACCCGCGACGACGTCCC                   CCGGGCCGTACGCACCCTCGCCGCCGCGTTCGCCGACTACCCCGCCACGC               GCCACACCGTCGACCCGGACCGCCACATCGAGCGGGTCACCGAGCTGCAA               GAACTCTTCCTCACGCGCGTCGGGCTCGACATCGGCAAGGTGTGGGTCGC               GGACGACGGCGCCGCGGTGGCGGTCTGGACCACGCCGGAGAGCGTCGAAG               CGGGGGCGGTGTTCGCCGAGATCGGCCCGCGCATGGCCGAGTTGAGCGGT               TCCCGGCTGGCCGCGCAGCAACAGATGGAAGGCCTCCTGGCGCCGCACCG               GCCCAAGGAGCCCGCGTGGTTCCTGGCCACCGTCGGCGTCTCGCCCGACC               ACCAGGGCAAGGGTCTGGGCAGCGCCGTCGTGCTCCCCGGAGTGGAGGCG               GCCGAGCGCGCCGGGGTGCCCGCCTTCCTGGAGACCTCCGCGCCCCGCAA               CCTCCCCTTCTACGAGCGGCTCGGCTTCACCGTCACCGCCGACGTCGAGT               GCCCGAAGGACCGCGCGACCTGGTGCATGACCCGCAAGCCCGGTGCCTGA          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 GCGGTGTTCGCCGAGAT 
                 (SEQ ID NO.: 55) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 GAGGCCTTCCATCTGTTGCT 
                 (SEQ ID NO.: 56) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 GCGGTGTTCGCCGAGAT 
                 (SEQ ID NO.: 57) 
               
            
           
         
       
     
      RIP7-rtTA  
                          (SEQ ID NO.: 58)                         ATGTCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCT                   TAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGC               TAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCT               TTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTT               TTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTA               AAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACAT               TTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATT               AGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCAC               TCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAG               CATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCC               GCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGC               CAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAA               CTTAAATGTGAAAGTGGGTCCGCGTACAGCCGCGCGCGTACGAAAAACAA               TTACGGGTCTACCATCGAGGGCCTGCTCGATCTCCCGGACGACGACGCCC               CCGAAGAGGCGGGGCTGGCGGCTCCGCGCCTGTCCTTTCTCCCCGCGGGA               CACACGCGCAGACTGTCGACGGCCCCCCCGACCGATGTCAGCCTGGGGGA               CGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGC               TAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCG               GGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGA               CTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACG               GTGGGTAG          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 TGCCAACAAGGTTTTTCACTAGAGA 
                 (SEQ ID NO.: 59) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CTCTTGATCTTCCAATACGCAACCTA 
                 (SEQ ID NO.: 60) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CCACAGCGCTGAGTGC 
                 (SEQ ID NO.: 61) 
               
            
           
         
       
     
      recombination—The process by which offspring derive a combination of genes different from that of either parent. In higher organisms, this can occur by crossing over.  
      recombinant DNA—A combination of DNA molecules of different origin that are joined using recombinant DNA technologies.  
      RNA—on of the two main types of nucleic acid, consisting of a long, unbranched macromolecule formed from ribonucleotides, the 3′-phosphate group of each constituent ribonucleotide (except the last) being joined in 3′,5′-phosphodiester linkage to the 5′-hydroxyl group on each ribose moiety renders these phosphodiester bonds susceptible to hydrolytic attack by alkali, in contrast to those of DNA. The RNA chain has polarity, with one 5′ end and on 3′ end. Two purines, adenine and guanine, and two pyrimidines, cytosine and uracil, are the major bases usually present. In addition, minor bases may occur; transfer RNA, however, contains unusual bases in relatively large amounts. The sequence of bases carries information, whereas the sugar and phosphate groups play a structural role. RNA is fundamental to protein biosynthesis in all living cells.  Oxford Dictionary of Biochemistry and Molecular Biology;  p. 577.  
      screening reference—are probes that are run on every sample submitted to screen laboratory. The probe is one that is found in every mouse, mutant or not.  
      Six-2 WT  
                              GGGTGAGGCTGTTGCGACGCCTCTTATTTAAAAAAAAAGGGAGGGGTGTCTCACAC   (SEQ ID NO. 62)           TTTTTCTCTTGAAGGCTCCTTCTGTCCCCCTCTTTTCCTTTCCTGAAAGGCACCCCCTT       AAACGGTCCTCCGCCTTCCCTTCTACTCCCTTCCTTCCCCACTTCGGTCCTCCTCTTTT       CTTCGAGGGCCCCCACCCAGCCCCCTCCTTCGGGGTCCTCCTCCTCCTCTGCTCTTTG       GGCGTCCGCCCCGTCAATCACCGCCGTCTCGGGGCCCCAGCCCGGCTCCTCTCCGCC       TCCCGGGCTCTGGGAGTGCCTGGGGCTCCCGTCTCGGCCAACCTCCGCTCTGTGCAG       AGCCGGGGCGATCTGTCAGCGGAGCTGGCCGAGGGGGGCGGGGGTGGGAGCCGCC       CGGGCCGCCGGGGCTCGGGTTACCGGTGACTGACAGCGTCTCCATGGCGAATAATTT       GACTCGACTATTGTCTGGCGCGGGCAGGCCCCGGGTCAGATAACCCGACCAATCAG       GGCGCGGGCCGCCGCGCCTCATGCCCGCTTAGAATAATATTATTAAAAAAGCTGCA       AGCGAGCTAGACGGGAGGGAGAGCGAACGAGCGAGGAGCCGGCGAGCGAGCGGCG       GGCGGGCGCGGAGCATGCGGAGCGGCGCCCCGGGCGGCCTCCGGGCTTGGGCGCGG       GCGAGGCGCGCGGGCGGCGGGGGCGCGGAGCTGCGCGGGGCCGGCGGCGGGAGCG       AGGACGGATCGTTGTGACTCAGGAGTCGCTCGGGAGCCGGCGCCTGGCCAGGGGGC       CCCGCCCGCCTGTCGGCCGGCCGGGGCCGGCGGGGAGGCGCCCATGCGGGGCCGCG       AAGCGCGGTGAGGGCGCGCGCGGGCGGGCGGGCGCGCAGCCGCCACCATGTCCATG       CTGCCCACCTTCGGCTTCACGCAGGAGCAAGTGGCGTGCGTGTGCGAGGTGCTGCA       GCAGGGCGGCAACATCGAGCGGCTGGGTCGCTTCCTGTGGTCGCTGCCCGCCTGCG       AGCACCTCCACAAGAATGAAAGCGTGCTCAAGGCCAAGGCCGTGGTGGCCTTCCAC       CGGGGCAACTTCCGCGAGCTCTACAAAATCCTGGAGAGCCACCAGTTCTCGCCGCA       CAACCACGCCA          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 GGGTTACCGGTGACTGACA 
                 (SEQ ID NO. 63) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CCCGCGCCAGACAATAGT 
                 (SEQ ID NO. 64) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CCATGGCGAATAATTT 
                 (SEQ ID NO. 65) 
               
            
           
         
       
     
      strain—a group of organisms bred for a genotype (at least one designated genetic sequence).  
      strain controls—are biomatter samples submitted by a remote user  1 . Strain controls are controls positive and negative sent to the screen laboratory as the remote user that discloses the genotype.  
      TetAKT1  
                              ATGAACGACGTAGCCATTGTGAAGGAGGGCTGGCTGCACAAACGAGGGGAATATAT   (SEQ ID NO.: 66)           TAAAACCTGGCGGCCACGCTACTTCCTCCTCAAGAACGATGGCACCTTTATTGGCTA       CAAGGAACGGCCTCAGGATGTGGATCAGCGAGAGTCCCCACTCAACAACTTCTCAG       TGGCACAATGCCAGCTGATGAAGACAGAGCGGCCAAGGCCCAACACCTTTATCATC       CGCTGCCTGCAGTGGACCACAGTCATTGAGCGCACCTTCCATGTGGAAACGCCTGAG       GAGCGGGAAGAATGGGCCACCGCCATTCAGACTGTGGCCGATGGACTCAAGAGGCA       GGAAGAAGAGACGATGGACTTCCGATCAGGCTCACCCAGTGACAACTCAGGGGCTG       AAGAGATGGAGGTGTCCCTGGCCAAGCCCAAGCACCGTGTGACCATGAACGAGTT       GAGTACCTGAAACTACTGGGCAAGGGCACCTTTGGGAAAGTGATTCTGGTGAAAGA       GAAGGCCACAGGCCGCTACTATGCCATGAAGATCCTCAAGAAGGAGGTCATCGTCG       CCAAGGATGAGGTTGCCCACACGCTTACTGAGAACCGTGTCCTGCAGAACTCTAGG       CATCCCTTCCTTACGGCCCTCAAGTACTCATTCCAGACCCACGACCGCCTCTGCTTTG       TCATGGAGTATGCCAACGGGGGCGAGCTCTTCTTCCACCTGTCTCGAGAGCGCGTGT       TCTCCGAGGACCGGGCCCGCTTCTATGGTGCGGAGATTGTGTCTGCCCTGGACTACT       TGCACTCCGAGAAGAACGTGGTGTACCGGGACCTGAAGCTGGAGAACCTCATGCTG       GACAAGGACGGGCACATCAAGATAACGGACTTCGGGCTGTGCAAGGAGGGGATCA       AGGATGGTGCCACTATGAAGACATTCTGCGGAACGCCGGAGTACCTGGCCCCTGAG       GTGCTGGAGGACAACGACTACGGCCGTGCAGTGGACTGGTGGGGGCTGGGCGTGGT       CATGTATGAGATGATGTGTGGCCGCCTGCCCTTCTACAACCAGGACCACGAGAAGCT       GTTCGAGCTGATCCTCATGGAGGAGATCCGCTTCCCGCGCACACTCGGCCCTGAGGC       CAAGTCCCTGCTCTCCGGGCTGCTCAAGAAGGACCCTACACAGAGGCTCGGTGGGG       GCTCTGAGGATGCCAAGGAGATCATGCAGCACCGGTTCTTTGCCAACATCGTGTGGC       AGGATGTGTATGAGAAGAAGCTGAGCCCACCTTTCAAGCCCCAGGTCACCTCTGAG       ACTGACACCAGGTATTTCGATGAGGAGTTCACAGCTCAGATGATCACCATCACGCCG       CCTGATCAAGATGACAGCATGGAGTGTGTGGACAGTGAGCGGAGGCCGCACTTCCC       CCAGTTCTCCTACTCAGCCAGTGGCACAGCCTGA          
 
     
       
         
           
               
               
               
               
            
               
                   
               
               
                 Forward Primer: 
                 GGAACGCCGGAGTACCT 
                 (SEQ ID NO.: 67) 
                   
               
               
                   
               
               
                 Reverse Primer: 
                 ACTGCACGGCCGTAGTC 
                 (SEQ ID NO.: 68) 
               
               
                   
               
               
                 Probe: 
                 CTGAGGTGCTGGAGGACA 
                 (SEQ ID NO.: 69) 
               
            
           
         
       
     
      Tetp27KIP  
                              CCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAG   (SEQ ID NO.: 70)           CTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGA       TGCCACCCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCG       TGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCT       ACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTAC       GTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGA       GGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACT       TCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCAC       AACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGAT       CCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACA       CCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGT       CCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTC       GTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAAG          
 
     
       
         
           
               
               
               
               
            
               
                   
                   
               
               
                   
                 Forward Primer: 
                   
                   
               
               
                   
                 CGTCGTCCTTGAAGAAGATGGT 
                 (SEQ ID NO.: 71) 
               
               
                   
                   
               
               
                   
                 Reverse Primer: 
               
               
                   
                 CACATGAAGCAGCACGACTT 
                 (SEQ ID NO.: 72) 
               
               
                   
                   
               
               
                   
                 Probe: 
               
               
                   
                 CATGCCCGAAGGCTAC 
                 (SEQ ID NO.: 73) 
               
            
           
         
       
     
      transgene—the foreign gene or DNA.  
      transgenic—this term describes an organism that has had genes from an organism or additional elements of it our sequence put into its genome through recombinant DNA techniques. These organisms are usually made by microinjection of DNA in the pronucleus of fertilized eggs, with the DNA integrating at random.  
      transgenic line—a transgenic mouse or organism strain in which the transgene is stably integrated into the germline and therefore inherited in Mendelian fashion by succeeding generation.  
      web site—a computer system that serves informational content over a network using the standard protocol of the World Wide Web. A web site corresponds to a particular Internet domain name such as TransnetYX.com.  
      wild type—the phenotype that is characteristic of most of the members of a species occurring naturally and contrasting with the phenotype of a mutant.  
      zygosity—This term reflect the genetic makeup of an individual. When identical alleles exist at a loci it is said to be homozygous; when alleles are different the alleles are said to be heterozygous.  
     2. OVERVIEW OF THE SYSTEMS COMPONENTS AND OPERATIONS  
      The present invention provides methods for genotype screening. More specifically, the present application relates to a method to rapidly screen biological samples for at least one designated genetic sequence. Various aspects of genotype screening involve: sample collection, lysing of the biological sample, isolation of purified genomic nucleic acid and nucleic acid screening. Additionally, the method operating according to the features described herein can provide screening results to a remote user  1  from the screening laboratory  20  within 24 hours of receiving the biological samples.  
      In order to screen for a designated genetic sequence, that sequence must first be determined or identified. Only when the designated sequence is known can a test be devised to search for its existence in the biological samples provided by the remote user  1  to the screening laboratory  20 .  
      There are a variety of ways the designated genetic sequence can be acquired by the remote user  1  or by the screening laboratory  20 . For example, if the sequence of bases that makeup the designated genetic sequence is known by the remote user  1 , the sequence can be directly communicated to the screening laboratory  20  via an electronic link, such as any of the electronic communication links identified herein, and particularly the communication links extending between the remote user&#39;s computer and the screening laboratory  20 .  
      The remote user  1  can indirectly communicate the designated genetic sequence to the screening laboratory  20  by communicating a publication, journal article, a gene name, a sequence name, a line or strain name (if the designated genetic sequence is found in animals of that line or strain), or the name of a mutation having the designated genetic sequence to the screening laboratory  20 . Alternatively, the remote user  1  can communicate to the screening laboratory  20  the sequence of a primer set or probe that corresponds to a target genetic sequence of the designated genetic sequence. These primer sets or probes will have previously been created or defined to indicate the presence of the designated genetic sequence.  
      The indirect references may provide the entire sequence. Alternatively, the screening laboratory  20  may take the information from the references or from the remote user  1  and use it to search public genetic databases such as The National Center for Biotechnology Information (NCBI), Ensembl, or The Wellcome Trust Sanger Institute database. The screening laboratory  20  can also search proprietary databases, such as the database provided by Celera Bioscience (Rockville, Md.).  
      Another indirect method that may be used to acquire or identify the designated genetic sequence is to use a third party who has specific knowledge of the sequence. For example, the screening laboratory  20  can receive the name of a transgenic animal line or strain from the remote user  1 , then contact the company that engineers that line or strain. The company can then transmit the sequence of bases that constitute the particular genetic sequence corresponding to that line or strain back to the screening laboratory  20 . These companies include such firms as Lexicon Genetics (Woodland, Tex.) or Charles River Laboratories (Wilmington, Mass.). Even further, individual researchers who have developed the line or strain, or who work with the same line or strain at another laboratory may provide the designated genetic sequence, the primer sets or the probes necessary to identify the designated genetic sequence.  
      If the designated genetic sequence is not known by the remote user  1  or third party and is not found in any public or private database, the screening laboratory  20  may use scientific methods. If the remote user  1  has a working genotyping assay, and they are performing PCR and separating fragments in a gel, the appropriate bands can be cut from the gel, purified and sequenced to determine the sequence of bases in that band. The company sequencing the bands can directly communicate the base sequence to the screening laboratory  20  or to the remote user  1 , who in turn can communicate the base sequence to the screening laboratory  20 .  
      Once identity of the designated genetic sequence is acquired by the screening laboratory  20  (and assuming a probe or primer set has yet to be designed), the screening laboratory  20  must then select a target genetic sequence of the designated genetic sequence for which a primer set and/or probe can be constructed. In the preferred embodiment, the sequence of the primer set and probe is determined using software such as Primer Express® (Applied Bio Systems). The target genetic sequence may be directly selected from the designated genetic sequence by the screening laboratory  20 . Once selected, the base sequence corresponding to the target genetic sequence is communicated to an oligonucleotide vendor, who manufactures the probe and primer sets and transmits them to the screening laboratory  20 .  
      The screening laboratory  20  preferably keeps a supply of probes and primer sets on hand so each future request by the remote user need not require special production of probes and primer sets.  
      Alternatively, a special probe or primer set may be required. In that situation, the screening laboratory  20  may not select the target genetic sequence itself, but may communicate to a third party specific areas in the designated genetic sequence that are important for mutation detection. The third party is typically an oligonucleotide vendor, who in turn will select the target genetic sequence, manufacture the probes and primer sets, and send the probes and primer sets to the screening laboratory  20 .  
      To effectively genotype these nontransgenic samples, additional bioinformatics are needed from the remote user  1 . Specifically, the screening laboratory  20  requests that the remote user  1  provide both the base sequence of the designated genetic sequence of the mutation as well as the DNA sequence of the endogenous location. The endogenous DNA sequence is disrupted if a mutation has occurred. Once the precise sequence data is acquired, two primer-probe sets are designed. The first primer-probe set determines if the sequence of the mutation is present, irrespective of the number of times it is present. The second primer-probe set determines if the endogenous DNA sequence is present. It is these two primer-probe sets that the oligonucleotide vendor designs and transmits to the screening laboratory  20 .  
      With respect to human genotyping, a remote user  1  can contact the screening laboratory  20  and provide information for a human mutation or suspected endogenous condition of interest. This information may include the remote user&#39;s interest in wanting to know if the sample is from a human or a mouse and if it is from a human what gender is the sample. The screening laboratory  20  can acquire primers and probe that can distinguish between humans and mice. This is accomplished by identifying areas of genetic sequence in the mouse genome that are not homologous with the genetic sequence in the  Homo sapiens  genome. With no input from the remote user  1 , the screening laboratory  20  can query a database such as Ensembl that would discriminate between the sex chromosomes in humans (X and Y). This query would yield sequence data for the Y chromosome, which is the designated genetic sequence. The screening laboratory  20  can take the designated genetic sequence, or portion thereof, and send it to a vendor indicating where to build the primer set and probe as to be informative for screening. Moreover, where there are a large number of nucleotides that are unique on the human Y chromosome, the screening laboratory  20  may send the sequence of bases to the vendor and have them build primer sets and probe anywhere inside the sequence. The remote user  1 &#39;s Internet web-based account will have a field populated that represents these reagents with an identifier such as the genetic line identification  84 . The remote user  1  will use the identifier (strain name or profile name) to indicate that these specific reagents are to be used on subsequent samples.  
      Similarly, if the remote user  1  requires SNP genotyping a remote user  1  can contact the screening laboratory  20  and provide a literature reference of the mutation which discloses the mutation name. A mutation name query of the Mouse Genome Informatics website yields links to different databases such as Ensenbl and National Center for Biotechnology Information that provides sequence data. This sequence data is the designated genetic sequence. Knowing the endogenous nucleotide and the mutant nucleotide, the screening laboratory  20  can take the designated genetic sequence, or portion thereof, and send it to a vendor indicating specifically where to build the primers and probes as to be informative for screening. For example, if the designated genetic sequence is 500 nucleotides in length, the screening laboratory  20  may indicate to the reagent vendor to build a SNP assay targeting the 239 th  nucleotide. The reagent vendor will then supply to the screening laboratory  20 , the primers and probes to specifically discriminate between a nucleotide change at the 239 th  position of the designated genetic sequence.  
      The remote user  1 &#39;s Internet web-based account will have a field populated that represents these reagents with an identifier such as a name or number, or what is commonly referred to as the genetic line identification  84 . The remote user  1  will use the genetic line identification  84  to indicate that these specific reagents are to be used on subsequent samples.  
      The probes and primer sets, if they are new and have not before been tested against a sample containing the designated genetic sequence, must then be tested, preferably by the screening laboratory  20 . To do this, the screening laboratory  20  preferably receives both a positive and a negative strain control samples from the remote user  1  and tests them against the probes and primer sets to confirm that they can be used successfully to determine whether the designated genetic sequence can be detected. These controls include one positive and one negative control for each mutation found in the strain of interest.  
      If the designated genetic sequence can be detected using the probes and primer sets, the screening laboratory  20  updates the website and the order management software to provide the remote user  1  with a web-based selection for sample testing using those tested probes and primer sets. These selections among which the remote user  1  can select are one of the screening parameter selections identified below.  
      Alternatively, for example, if the remote user  1  or other third party communicates to the screening laboratory  20  that a particular probe or primer set has already been tested and is known to work, or if the screening laboratory  20  has already designed a probe and primer set for the designated genetic sequence (which is commonly the case for often-used strains or lines of transgenic animals) the screening laboratory  20  can immediately add a selection to the website and does not need to test controls with the probes and primer sets.  
      The strain controls are used to tell LIMS  24  a signal magnitude that is then associated with a positive or negative sample. In one case, the remote user  1  may send these controls together with the samples to be tested to the screening laboratory  20  in a single shipment. Alternatively, the controls may be sent separately from the samples to be tested.  
      The screening laboratory  20  tests the strain controls using the process described herein for testing samples. At the end of this testing process, the signal values for the strain controls are recorded into LIMS  24 . The magnitude of the signal provided by the positive control indicates the expected signal level for subsequently tested samples having the designated genetic sequence. The magnitude of the signal provided by the negative control indicating the expected signal level for subsequently tested samples that do not have the designate genetic sequence.  
      The computer at the screening laboratory  20  is configured to compare the test results (i.e. signal levels) for every sample that it subsequently tests for that designated genetic sequence with these multiple control signal levels and, based on that determination, to decide whether that sample has or does not have the designated genetic sequence. Positive and negative strain controls for a line therefore do not need to be resubmitted for each subsequent order but can be referenced by the screening laboratory  20  computer when later samples are tested for the same designated genetic sequence.  
      For transgenic zygosity genotyping, additional controls (not just a positive and a negative) are required to indicate each possible variation such as: a homozygous control, a heterozygous control and a wild type control.  
      Upon receipt of the primers and probe from a vendor, the sample, if available, will be screened using these reagents. Once a determination is made that there is discrimination between different genetic conditions, then the reagents will be placed in the inventory. Additionally, the screening laboratory  20  will populate a data field on the order management system, allowing the remote user  1  to select this primer sets and probe combination(s) for subsequent samples. This data filed will be populated with an indicator such as a mutation name, strain name or genetic line identification that will represent these reagents or combination of reagents that will be used in subsequent samples of this strain. This allows the remote user  1  to select the indicator of the reagents and prevents the need to transfer genetic information with each order.  
       FIGS. 1-3  present an overview of certain features of the present invention. The present invention allows a remote user  1  with access to a computer  5  to order genotype screening of samples they submit to screening laboratory  20 . Using the Internet or other communication link  7 , the remote user  1  sends an access request from the remote user&#39;s computer  5  to a screening laboratory  20  computer  9  via an electronic communication link  7 , such as the Internet. The screening laboratory  20  website  19  will transmit an access enabling response to the remote user  1  via electronic communication link  7 . This response includes three distinct sections. The three sections are Account Registration  21 , Survey of Work  23  and Sample Identification and Designation  25  ( FIG. 3 ).  
      Now referring to  FIG. 2 , a remote user  1  can access screening laboratory  20  website  19  via communication link  7 . The website  19  can be housed by an order manager  22 . An order manager is a software-based order management system. In the preferred embodiment the order manager  22  is an order management system developed by “Big Fish”, a software development company in Memphis, Tenn. The order manager  22  functions to manage the placement of the order. The order received from the remote user  1  is transmitted to website  19 , which reports the order to order manager  22 . Manager  22  is in electronic communication via link  7  with screening laboratory  20  computer  9 . Screening laboratory  20  computer  9  includes LIMS  24 , which is communicatively coupled to a process controller  26 .  
      LIMS  24  is the generic name for laboratory information management system software. The function of LIMS  24  is to be a repository for data, to control automation of a laboratory, to track samples, to chart work flow, and to provide electronic data capture. LIMS  24  can also, in another embodiment, be in direct communication with the remote user  1  via an electronic communications link  7 . Any standard laboratory information management system software can configured to be used to provide these functions. Alternatively, a standard relational database management system such as Oracle (Oracle Corp., Redwood Shores, Calif.) or SQL Server (Microsoft Corp., Redmond, Wash.) either alone or in combination with a standard LIMS system can be used. In the preferred embodiment, the Nautilus® program (Thermo LabSystems, a business of Thermo Electron Corporation, Beverly, Mass.) is used.  
      The process controller  26  is communicatively coupled to the workstation  14 . The process controller provides commands to any portions of the workstation  14  that are amenable to automation. For example, process controller  26  directs the delivery of the probes and primers to the Screening Station  95 . The workstation  14  is communicatively linked  28  to LIMS  24 . In this way, the workstation  14  can provide data to LIMS  24  for the formulation of the outcome report  249 , and then, via link  7  to the order manager  22  or remote user  1 . In an alternative embodiment, remote user  1  at remote user computer  5  can be linked  7  to the screening laboratory  20  by a direct phone line, cable or satellite connection.  
      Now referring to  FIG. 4 , the user&#39;s Account Registration section  21  begins with logging into the system  30 . A remote user  1  accesses an existing account by entering an account identification  31 , which is, for example, an e-mail address. The user will then enter a password  37 . If a valid password is entered, the user can place a new order  39 . Alternatively, the user can check an order status  41  by providing an order number  43  and can proceed to order tracking  45 . Alternatively, a new account  47  can be opened by providing an institution name, principal investigator, address, phone number, fax number, electronic mail address, billing information, and other authorized user names  49 . The user can enter a password  51 , confirm the password  53  and enter this billing information  55 .  
      Now referring to  FIGS. 5-6 , once the remote user  1  submits the Survey of Work section  23  the remote user  1  will be presented with the Sample Identification and Designation section  25 . In this section, the user (among other things) identifies where he will place each sample to be tested in an actual (physical) container  2  ( FIG. 1 ) by associating each sample with a corresponding well of a virtual 96 well container displayed on the computer screen of computer  5  as described below. The Sample Identification and Designation section  25  includes 96 well container locations. The remote user  1  designates which sample was or will be placed into each well. If the remote user  1  has more than 96 samples, subsequent 96 source well containers and designations are available. With respect to  FIG. 6 , a 96 well source well container  2  having a barcode accession number  3  ( FIG. 1 ) will be shown ( FIG. 6 ) oriented in the longitudinal direction having an X axis labeled “A” to “H” (at  80 ) and a Y axis labeled “1” to “12” (at  81 ). The X and Y axes designate a well position such as “A1”.  
       FIGS. 5 and 6  together illustrate the Survey of Work section  23  and the Sample Identification and Designation Section  25 . Referring now to  FIG. 5 , the remote user  1  is asked to provide: source well container  2  accession number  82 , which the remote user  1  gets from the accession number  3  on the physical source well container  2  at his facility ( FIG. 1 ) that he intends to fill (or has filled) with the samples, number of lines  83 , genetic line identification  84 , number of samples  85 , and well location  88 . The remote user  1  is also asked for any internal sample identification number  91 .  
      For genotyping (i.e. screening to determine the presence of a designated genetic sequence) the positive strain control and the negative strain control samples are designated and deposited in wells of a microwell container. The remote user  1  indicates that a sample is a control sample at  89 . This assumes, of course, that the strain controls were not earlier provided to the screening laboratory  20  as described above. If a control is deposited in source well container  2 , remote user  1  can also designate the zygosity, mosaic nature and copy number of the sample.  
      At this point, the remote user has completed the Survey of Work section  23  and the Sample Designation section  25  of  FIGS. 5-6  and is ready to transmit the screening parameter selections gathered in those sections to website  19  and thence to screening laboratory  20  computer  9 .  
      Now referring to  FIGS. 1 and 2 , the remote user  1  transmits his or her order including the completed screening parameter selections to the screening laboratory  20  via link  7  such as the Internet or a direct line. The remote user  1  can transmit the selected screening parameter selections to LIMS  24  in screening laboratory  20  via electronic communications link  7 . This link  7  can be direct or indirect. In the indirect route, the screening parameters are first transmitted to web site  19 , wherein order manager  22  receives the order and then provides LIMS  24  with the screening parameter selections.  
      In a particularly preferred embodiment of the system described in the foregoing paragraphs, remote user  1  at computer  5  transmits a request for a home web page served by screening laboratory  20  web site  19  via the electronic communication link  7 . Web site  19 , in turn, serves a home web page to computer  5  that includes information identifying the source of the web page and including a login button. Remote user  1  at computer  5  clicks on the login button displayed on his computer screen, transmitting a signal to web site  19  requesting access to the web site. This request is transmitted over communications link  7  to web site  19 , which responds with a second web page having fields for the entry of an account identifier (in the preferred embodiment an e-mail address), and a password. Remote user  1  enters the remote user  1  e-mail address and password, and transmits this information to web site  19  to gain access to the web site. Web site  19  receives this access request and compares the account identifier and password against its database of pre-existing accounts in the order manager  22  to determine whether the user is permitted to access the web site  19 . If so, computer order manager  22  serves up a further web page, called an order manager web page, which includes several user selectable choices including an “order status” button for tracking previous orders and results (if any have been received), a “supply request” button for requesting supplies, and an “order” button for ordering additional tests.  
      To order genetic testing, user  1  clicks on the “order” button displayed on the screen of computer  5 . Computer  5  transmits the user  1  request to web site  19 . Web site  19  receives this request, and transmits a first ordering web page to computer  5 . Computer  5 , in turn, displays several fields on its computer screen, including several data entry widgets. The first of these widgets is list box including two selectable entries for requesting the speed of service. In the preferred embodiment there are two speeds of service: 24-hour service and 72 hour service. The second of these widgets is a list box providing several entries, each entry in the box corresponding to a strain for which the sample is to be tested. The third widget is a text box for entering the number of samples of the selected strain to be tested. The fourth widget is a text box for entering the accession number (typically a bar code number) of the source well container  2  in which the samples are to be placed for shipping to the screening laboratory  20 .  
      The remote user  1  types in the number of samples to be tested. In this embodiment the samples are taken from transgenic animals, each sample typically corresponding to one animal to be tested. Typically several animals are tested to determine if they received the transgenic gene from their parents. Each strain of animal is defined by one or more designated genetic sequence. Thus, by designating the strain for which the samples are to be tested, the remote user  1  selects the one or more designated genetic sequences associated with that sequence. In the preferred embodiment, the remote user  1  can also select or deselect each individual probe and primer set that is used to screen for the designated sequences in the strain or line of the biological sample.  
      Once the remote user  1  has entered the number of samples to be tested, he or she then enters the name of the strain that the samples are to be tested for. Again, by selecting a strain the remote user  1  indicates the designated genetic sequence for which the samples are to be tested, since each strain is bred to have that sequence.  
      Once remote user  1  has selected the speed of service, the strain to be tested, and the number of samples to be tested for that strain, he enters the accession number from the source well container  2  and clicks on a button on the first ordering web page for recording this first group of samples to be tested. Computer  5 , in turn, generates a revised first ordering web page, the revised page including a table entry in a table on the revised web page listing the first group of samples in tabular form, wherein each row in the table corresponds to one group of samples to be tested, identifying that group of samples by the strains for which that group of samples is to be tested, and the number of samples in that group.  
      This process of creating a new group of samples and identifying them by the strain for which they&#39;ll be tested, and the number of the samples, can be continued as many times as necessary until all the samples to be tested are identified in the table.  
      Once all of the groups of samples have been entered and listed in the table on the revised first ordering web page, the operator then selects a button identified “next” and moves to the next stage in the ordering process. Computer  5  transmits this request to web site  19 , which generates a graphical image of a 96 source well container, appearing on the screen of computer  5  identical to the corresponding 96 source well container  2  that the remote user  1  is filling/has filled with samples, and transmits that image embedded in a second web page back to computer  5  for display. The second web page includes a graphical representation of a 96 well plate, in a top view, showing the two dimensional array of all 96 wells in which the remote user  1  is to place the samples identified previously. Web site  19  calculates the respective positions of each group of samples in the well container  2 . Each group is shown in the graphical representation of the well plate in a different color. All the wells in a group are shaded with the color associated with that group.  
      Samples of the same color from the same group are grouped together thus producing several different contiguous groups of wells, each group of wells have the same color different from the color of the adjacent groups.  
      The images of the wells in the web page are displayed on the computer with an initial shading to indicate that they have not been identified to a particular animal from which the sample in each well will be taken. In the preferred embodiment, each well contains a sample, such as a tissue sample, taken from an individual animal. The purpose of the testing performed on the samples in the wells is to determine the genetic characteristics of the animal from which each sample was taken. In order to relate the test results performed on each sample back to the animal from which the sample was taken, the user must make a record of the animal source of each sample (i.e. the animal from which each sample was taken).  
      To uniquely identify each sample in each well with an associated animal, remote user  1  selects a button on the third ordering web page. This button signals computer  9  to generate an additional web page. This web page lists each well in the well plate that was previously identified as containing a sample. Thus, if the first group of samples were 13 in number, there would be 13 entries listed in the additional web page. The web page itself is arranged as a single column of entries. Each entry in the column of entries includes a well identifier (called well location  88 , above), which is a string of alphanumeric characters that uniquely identifies one well of source well container  2 . A preferred well identifier for the 96 well plate is an alphabetic character followed by a numeric character. A text box is adjacent to each well identifier on the additional web page. To uniquely identify each sample in the source well container  2 , the user enters alphanumeric characters in the text box that are uniquely associated with each sample. This identifier is typically a short string of consecutive alphabet or numeric characters, a practice commonly used by research facilities to identify individual animals used for testing.  
      Animals in a particular group of animals having (presumed) common genetic characteristics will typically be identified by tattoos, tags, or other permanent means by consecutive or sequential numbers, characters, or combinations of numbers and characters (for example “A1”, “A2”, “A3”, or “101”, “102”, 103”, or “AA”, AB”, “AC”, etc.). In a preferred embodiment, user  1  enters each animal number into the text box as a sample ID  91 . Animals may also be identified by a unique combination of disfigurements such as cutting or cropping toes, tails or ears that can also be approximated to a progressive alphanumeric sequence.  
      To assist the remote user  1  in entering the sample ID  91  into each of the text boxes in the additional web page, a button is provided to automatically fill several consecutive text boxes based upon the alphanumeric characters typed into a few text boxes from the group. For example, if the user types in “B7” in the first text box of a group, then types in “B8” in the second text box of a group, computer  5  is configured to automatically generate consecutive alphanumeric strings to fill the remaining text boxes of the group based upon these two manually typed-in entries. In this case, computer  5  would automatically generate the alphanumeric strings “B9”, “B10”, “B11”, etc. and insert these characters sequentially into the remaining text boxes of the group in the additional web page. This process can be repeated for each subsequent group shown on the additional web page. Alternatively, the computer can be configured to automatically generate alphanumeric characters for all the groups at once and to fill the text boxes of all the groups all at once. Once the user has finished identifying all of the groups of samples and filling out all of the sample ID&#39;s  91  in the text boxes on the screen of computer  5 , he clicks on a button labeled “next”. Computer  5  transmits this request to website  19 , which responsively generates another web page in which the user  1  enters shipping and tracking information. This page, called the order confirmation page, includes a text box for entering a character string. This character string provides access to a web-based shipment tracking system of a commercial shipping company. In the preferred embodiment, the character string is a tracking number used by the shipping company to track the samples from the remote user  1  to the screening laboratory  20 . In the preferred embodiment, the tracking number is provided to the user together with the source well container  2  and the packaging materials in which the user places the source well container  2  for shipment to the screening lab  20 .  
      The order confirmation page also includes an invoice that lists the different tests requested by the operator in the foregoing steps on the screen of computer  5 . Each test or group of tests is displayed on the screen adjacent to the price or prices for those tests. A total price of all the tests is displayed as well.  
      The order confirmation page has a second text box in which the remote user  1  can type the expected shipping date. The expected shipping date is the date on which remote user  1  intends to give the samples in their packaging materials to the delivery service associated with the tracking number. By providing the anticipated shipping date to the website  19  and then to the screening laboratory  20 , personnel at the screening laboratory  20  can anticipate the arrival of each shipment and prepare for its arrival by pre-ordering reagents, probes and primer sets required for testing the samples in advance.  
      Once the operator has entered the tracking number and the expected shipping date, he clicks on a button labeled “confirm order”, which transmits the completed order, including the tracking number and expected shipping date to website  19  and order manager  22 , and thence to LIMS  24 .  
      In the preferred embodiment, once the order has been transmitted to the order manager  22 , the order generates two electronic messages, which will be sent to different locations. The first message is cross-referenced in LIMS  24  with a list of stocked probes. If the probe designated by the user is not stocked, an order message is sent to a supplier  11 , such as a contracted probe provider. This request can be transmitted from remote user  1  to screening laboratory  20  via any form of electronic communication, and then via a form of electronic communication  10  to suppliers&#39; computer  8 , or in the alternative, the order message can go from user  1  via any form of electronic communication link  12  to suppliers&#39; computer  8 . The supplier  11  creates the primer sets and probe based on the designated genetic sequence designated by the remote user  1  or the screening laboratory  20 . The made to order probe can be referred to as the target-binding probe. This supplier  11  will then barcode and overnight ship  13  the primer sets and target-binding probes  17  to the screening laboratory  20 . Once the primer sets and target-binding probes for each order for that day&#39;s screening are received by screening laboratory  20 , the barcodes on the primer sets and target-binding probes are scanned into LIMS  24 . The LIMS  24  records the date and time the primers and target-binding probes were received along with the quality control data provided from the probe provider.  
      In the preferred embodiment, the primer sets and target-binding probes are placed in workstation  14  and LIMS  24  will record the barcode of the probe and record its specific location on the deck of the workstation  14 , as will be discussed in more detail with respect to the Screening Station  95 . Additionally, the screening laboratory  20  and the LIMS  24  system correlates which target-binding probes will be used on which samples, as will be discussed in more detail with regard to the Screening Station  95 .  
      The second message, in the preferred embodiment, that is generated from the order placement of the remote user  1  insures that the remote user  1  has the proper supplies to package and ship their samples. This message, sent via link  12 , will define the barcode number of well container(s), shipping labels tracking number and amount of reagents needed for the user. In response to this message, supplier  11  will package  18  supplies for remote user  1  and ship  14 A the supplies back to remote user  1 .  
      Once the remote user  1  procures or receives these supplies, the remote user  1  places the appropriate samples into the source well containers  2  previously identified in the order sent to website  19 , order manager  22  and LIMS  24 . In other words, the remote user  1  fills each well of source well container  2  such that each well contains the same sample with the same sample ID  91  that the user previously identified in the order previously sent to website  19 . Alternatively, if the user already had sufficient supplies when the user placed the order the user need not wait for a source well container  2  to be sent by a supplier, but can fill the source well container  2  when the user creates the order, or even before the order is created. What is important is that the contents of the actual 96 source well container  2  that the user fills exactly matches the description of the samples and has the same accession number as the order the user previously sent to website  19 .  
      The samples can be obtained from prokaryotic or eukaryotic organisms. The samples may be a tissue, cells or biological fluid such as blood, lymph or semen sample from a mouse  8 A, but can also come from other animals (including humans), plants and viruses. In the preferred embodiment, mouse oral cavity swabs or anal cavity swabs provide a sample. Source well container  2  is a 96 well plate or the like that receives the sample in each well of the well plate. A sufficient amount of lysis reagent can be added to cover the sample. In one embodiment, the lysis reagent is added prior to transit to the screening laboratory  20 . Although, in the preferred embodiment the lysis reagent is added at the screening laboratory  20  at Lysing Station  92 .  
      A biological sample can be collected in a variety of ways to facilitate rapid screening. In one embodiment, the collection method involves swabbing the oral, nasal or anal cavity of an animal to be tested, such as a mouse, to collect cells for screening. In this collection method swab tips are removed by the remote user  1  and placed in individual wells of a multi-well container for transport to the screening laboratory  20 . Many different swab materials may be used including polyester, cotton, acrylamide, nylon and calcium alginate. In the preferred embodiment Microbrush® (Graftin, Wis.) swabs are used. A multi-well container as shown in  FIG. 1 , in the preferred embodiment, is a 96 microwell source well container  2  but can include other multi-well containers, such as Strip Racks, 24 well plates, 384 well plates and tube rack holders or the like. As described above with regard to  FIG. 6 , the remote user  1  operates computer  5  to enter a variety of data regarding the samples placed in the source well container. Once all of the samples in all of the wells have been identified in this manner, the remote user sends the source well container  2  containing a plurality of biological samples to a screening laboratory  20  for screening.  
      Now referring to  FIG. 20A and 20B , an apparatus to swab the subject and to facilitate placement of the swab into a source well container  2  is disclosed. A swab holder  300  with disposable swab  301  is shown. The swab  301  has a proximal and a distal end with respect to a swab holder  300 . The distal end of the swab  301  is made of a sufficient amount of flocking to collect a biological sample. The proximal end of the swab  301  has at least one annulus  305 . The function of the at least one annulus  305  is to secure the swab  301  to the swab holder  300  during swabbing of a subject. The swab holder  300  preferably includes an elastomeric, rigid plastic grip area, metal or the like on outer surface with metal, metallized plastic or the like main body. The body of the swab holder  300  is linear with respect to the swab  301  to facilitate collection of biomatter. A spring loaded plunger  306  has a release button  307  on opposite end from swab  301 . The action is like that of a retractable ball point pen but without the latch function.  
      The swab holder  300  has an internal section configured to retain at least one annulus of a swab  301 . In the preferred embodiment, the internal section  304  is deformable. This section can be elastomeric, serving as a swab grip, which receives and holds disposable swab  301  until released by the spring plunger  306 . The mounting end of the swab tip has at least one annulus  305  which, upon insertion into the swab grip, deforms or squeezes into the elastomer sufficiently to retain the swab  301  during its function. Although three annuli are shown in the  FIG. 20A , it would be possible for one elongated annulus to serve the purpose.  
      In the preferred embodiment, the swabs  301  are composed of a plastic material that measures approximately 1 inch long with a diameter of approximately 0.050 inches. The distal portion of the swab  301  is flocked with nylon fibers. Whereas, the proximal end of the swab  301  shaft is designed to fit into the swab holder  300 .  
      After the swab  301  is seated in the swab holder  300  the remaining portion of the swab  301  shaft and flocking are inserted into an orifice of a subject to collect biomatter. The swab  301  and/or swab holder  300  may be rotated to facilitate the collection of biomatter. Upon sufficient collection of the biomatter, a mechanism  307  is depressed on the swab holder  300 , such as a button that ejects the swab  301  from the distal end of the swab holder  300 . The ejector mechanism is then loaded with a new swab  301  and the process is repeated as many times as necessary.  
      In another embodiment of this invention, the biological sample is a blood sample collected by nicking the animal to be tested and blotting the blood on a filter paper. The blotted filter paper is placed in individual wells of source well container  2  by the remote user  1  and transported to the screening laboratory  20 . In both of these embodiments, the biological sample is disposed on an absorbent carrier.  
      Now referring to  FIG. 21 , the swab holder apparatus  300 , swab  301  and a source well container  2  can be packaged in a kit  310  and sent to a remote user  1 . The kit  310  does not need to be sterilized.  
      Referring now to  FIG. 1 , source well container  2  has an accession number  3  affixed to the side of the container. The accession number is used by LIMS  24  to track the source of source well container  2 . The remote user  1  places the appropriate samples into the well locations in source well container  2  that they had previously designated while placing their order in  FIG. 6 . The remote user  1  will add lysis reagent  4  to each well of the source well container  2 . The lysis reagent  4  should completely cover the samples. Once the samples and lysis reagent  4  are in the source well container  2  the remote user  1  places a seal on the top of the source well container  2  preventing samples from leaking. The remote user  1  then places a plastic lid on the seal for transportation. The remote user  1  then places the source well container  2  into an overnight delivery service package and shipped frozen  15 . The remote user  1  will then seal the package and ship  16  to screening laboratory  20 , and apply a barcode shipping label.  
      Now referring to  FIG. 7A -D, the preferred embodiment of the present invention is shown. In  FIG. 7A , the source well containers  2  arrive  101  at the screening laboratory  20 . The tracking number of the shipping label is read with a barcode reader  103 . If the shipping label is unreadable  105 , the tracking numbers are manually entered  107 . The scanning of the tracking number is received  104  in LIMS  24  and a received message is posted to the user&#39;s account as shown in tracking field. The source well container  2  are removed from the package and taken to a clean room  109 . The source well containers  2  contain the raw biological matter and in one embodiment lysis reagent. The source well containers  2  individual barcodes are scanned by the barcode reader  111  and recorded  106  in LIMS  24  as accession numbers. LIMS  24  can send  106  a probe order to supplier  11  through the order manager  22 . If the source well containers  2  individual barcodes are unable to be scanned  113 , the accession numbers are entered manually  115 . If the tracking number, accession number, user order and worklist properly correlate, LIMS  24  will activate (not shown) an active record number for the containers.  
      The source well containers  2  are loaded  116  into a transportation apparatus in a clean room. A transportation apparatus is any device that holds well containers and that can dock with the workstation. The transportation apparatus, in the preferred embodiment, includes several rigid trays stacked vertically in a housing unit that is mobile. This transportation apparatus can be moved between different automated stations, docked and the rigid trays can be removed in an automated fashion and processed on the deck of a workstation. Each rigid tray consists of nine locations for source well containers  2 . Each of these nine locations per tray has a unique barcode designating its specific location inside the trays of the transportation module.  
      Source well container  2  accession number  3  is scanned with a barcode reader and the bar-coded source well container  2  location in the transportation apparatus trays is scanned. The barcodes of source well containers  2  are married  117  in LIMS  24  with the unique barcode locations in the transportation apparatus trays for tracking purposes. LIMS  24  records and associates each well container to this location. Once the transportation apparatus is loaded with the source well containers  2 , the transportation apparatus is docked  119  into the laboratory workstation  14 .  
      LIMS  24  will generate a worksheet for laboratory personnel (not shown). The worksheet outlines the probes and primer sets that the operator will need to prepare or gather in order to test the latest samples. The LIMS  24  worklist will generate a single file. The file format may include, but is not limited to, ASCII, XML or HTML. The file will be written into a specified directory on the network drive. The name of the file will be unique and will correlate to a run number. The extension will be unique for worklist files.  
      In the configuration described above, a transportation apparatus includes a housing unit provided to support several trays, each tray having nine different locations for nine source well containers  2 . In an alternative embodiment, however, the housing unit can be eliminated. Instead, the source well containers  2  can be manually transported throughout the workstation in trays from functional station to functional station. In this system, operator at the laboratory loads source well containers into the trays after the source well containers  2  are received at the screening laboratory  20  and are scanned into LIMS  24  as described above for transportation to workstation  14 . Alternatively, source well containers  2  can be transported individually to workstation  14  and be placed in a tray or trays that are already located at workstation  14 .  
      We now refer to  FIG. 8 , which depicts one embodiment of the workstation  14 . Standard laboratory stations are logical groupings of laboratory operations. These groupings, however, do not necessarily refer to different physical stations. These logical groupings include: Lysing Station  92 , Automated Accessioning Station  93 , Isolation/Purification Station  94 , Screening Station  95  and Detection Station  96 , all of whom make up the workstation  14 . The Screening Station  95  can include other screening processes such as PCR. Lysing Station  92  is an alternative step provided to lyse the samples in containers  2  in the event user  1  does not choose to lyse the samples by adding a lysis reagent before sending them to laboratory  20 . The functions of the various logical stations are described below in connection with the steps shown in FIGS.  7 A-D. The following description provides the preferred embodiment, although one skilled in the art could elect to conduct these methods with varying degrees of automation as required.  
      As mentioned above, remote user  1  need not add a lysis reagent to the samples before shipping them to screening laboratory  20 . Instead, the samples may be shipped un-lysed (frozen) and may be lysed at laboratory  20  by piercing the cover  121  of the container  2  and treating each of the samples with a lysis reagent after docking the tray in the workstation  119  in the lysing station  92 . The samples are incubated  123  to produce a lysate containing cellular debris including at least a portion of intact genomic nucleic acid.  
      With respect to the swab sample collection method, the preferred embodiment is to have the swabs shipped without lysis solution. A sufficient amount of a lysis reagent, such as SV Lysis reagent or Nucleic Lysing Solution (Promega Corporation, Madison, Wis.) is added to each well of source well containers  2  to cover the swab tips at the screening laboratory. Swabs do not need to be incubated for three hours, however they are voretexed for ten minutes in the lysis solution.  
      With respect to the blood sample collection method, a sufficient amount of a lysis reagent, such as Nuclei Lysing Solution (Promega Corporation, Madison, Wis.) is added to each well of source well containers  2  to cover the filter paper after shipment. With respect to animal embryonic and stem cell screening, Nuclei Lysing Solution (Promega Corporation, Madison, Wis.) is added to each well containing the tissue. The source well container  2  is treated under conditions to facilitate rapid lysis of the biological sample. In the preferred embodiment, these conditions are heating at 55° C. for three hours.  
      The preferred method of performing the above lysing steps at Lysing Station  92  includes loading source well containers  2  into the tray  9206  and taking the rigid tray to Lysing Station  92  to be lysed. Lysing Station  92  includes a liquid handler  9220 , such as Genesis Tecan (Raleigh Durham, N.C.) or Multimeck Beckman (Indianapolis, Ind.). An example of a preferred Lysing Station  92  is shown in  FIG. 14 . It includes a frame  9202 , on which a deck  9204  is mounted to provide a horizontal working surface, which supports tray  9206 , which supports and positions up to nine source well containers  2 . A material handler  9214  is fixed to frame  9202  and extends upward and across the top surface of deck  9204 . A computer  9208  is coupled to material handler  9206  to direct the movement and operation of pipettes  9210 . A trough or reservoir  9212  is provided on deck  9204 , from which computer  9208  commands the material handler  9214  to aspirate lysis reagent into pipettes  9210  and to deposit the reagent into wells of container  2 .  
      The operator first carries a plurality of source well containers  2  and places them on deck  9204  in one of the nine positions on the rigid tray  9206  that support and orient source well containers  2  thereby docking them  119  into the workstation  14 . The operator then enters the number of wells that are filled with samples in each of the source well containers  2  into computer  9208  in combination with the location of that container with respect to tray  9206 .  
      Knowing the location of each source well container  2  in tray  9206 , and the number of wells that are filled with samples in each of these source well containers  2 , computer  9208  then directs material handler  9214  to move the pipettes  9210  to each source well container  2  in turn, piercing  121  the barrier sealing mechanism and filling each of the wells of source well containers  2  containing a sample with lysis reagent. By providing the location and the number of samples, computer  9208  is configured to fill only the wells containing samples with lysis reagent and to leave the empty wells empty of lysis reagent.  
      Once each of the sample-containing wells has been filled with lysis reagent, the operator moves the entire tray or trays  9206  containing the samples to an oven  9216  ( FIG. 15 ), where the samples are incubated  123  by heating for a period of about three hours at a temperature of 55° C. (described-above). Once the incubation process is complete, the operator moves source well containers  2  supported on the tray or trays  9206  to Automated Accessioning Station  93 .  
      An Automated Accessioning Station  93  provides a device to remove liquid from the source well container  2  to the primary master well container  6 . The primary master well container  6  is the container in which the nucleic acid is isolated. It is preferably a 384 well plate (Fisher Scientific #NC9134044). Any commercially available automated accessioning device can perform this function such as Genesis® Tecan (Raleigh-Durham, N.C.) or Multimeck® Beckman (Indianapolis, Ind.). These devices are referred to as liquid handlers. The source well containers  2  barcode accession numbers  3  are re-scanned  127 . This measurement will be recorded and posted  108  into the LIMS  24  database and reflected in the outcome report  249 . Additionally, LIMS  24  ensures  108  that source well containers  2  are consistent from transportation apparatus to the Automated Accessioning Station  93 . Error codes will be generated if a sufficient amount of raw testing material is not available. The liquid handler utilizes stainless steel, or the like, pipette tips that are washed between each sample transfer. Alternatively, disposable pipette tips may be used.  
      The nucleic acid lysate is transferred  129  to clean well containers, called primary master well containers  6 . Each of the containers  6  has a scannable accession number, preferably a barcode accession number, called “barcodes” or “accession numbers” below. The barcodes of the primary master well containers  6  are scanned  131  and LIMS  24  marries  102  the barcodes for the primary master well containers  6  to the scanned barcode accession numbers  3  of the source well plates  2 . The automated process accessioning continues until all of the day&#39;s pending samples are accessioned into the primary master well containers  6 . The preferred method of performing the above steps at Accessioning Station  93  includes taking the rigid tray  9206  and the source well containers  2  from the incubating oven  9216  back to the same liquid handler  9220  that performs the functions of Lysing Station  92 . This liquid handler  9220  is also preferably configured to function as Accessioning Station  93 .  
      Referring now to  FIG. 14 , the operator returns tray  9206  to liquid handler  9220  and places tray  9206  back on deck  9204  generally in the same location it was in when the lysis reagent was inserted into each well containing a sample.  
      Once in that location, the operator commands computer  9208  to fetch the work list from LIMS  24  and electronically stores it in the computer memory of process controller  26 . The work list includes the accession numbers of each source well container  2  that is in tray  9206 , together with the probe type that should be used for each well. The work list uniquely associates the location of the well, the accession number of source well container  2  from which the well is from, the probe type that is to be used with the sample in that source well container  2 , and the quantity of probe to be added to that sample.  
      Once computer  9208  fetches the work list, computer  9208  directs the operator to electronically scan  127  the accession numbers of all the source well containers  2  that are in rigid tray  9206  on deck  9204  of liquid handler  9220  using scanning device  9218  coupled to computer  9208 . Scanning device  9218  is preferably a glyph scanner, character scanner, bar code scanner, dot matrix scanner, or RFID tag scanner, depending upon the form of the accession identifier (typically a barcode accession number  3 ) on source well container  2 . Once source well containers  2  have been scanned  127 , computer  9208  transmits  108  the accession numbers  3  to process controller  26  and thence to LIMS  24 . Process controller  26  preferably includes an instrument database to which each of the computers of Lysing Station  92 , Automated Accessioning Station  93 , Isolation/Purification Station  94 , Screening Station  95  and Detection Station  96  transmit their data in order to maintain an ongoing record of the testing process and the location of materials and samples throughout that process. The database is preferably implemented using Microsoft&#39;s SQL Server, although any relational database (e.g. Oracle), may be used.  
      Computer  9208  then commands material handler  9206  to transfer  129  the contents of each well (i.e. lysate) in source well containers  2  to a corresponding well in the primary master well container  6  using pipettes  9210 . Computer  9208  directs the operator to scan  131  the accession numbers on the primary master well container  6 . Like the accession number on source well containers  2 , the accession number on the primary master well container  6  may be any electronically scannable indicia or device. Computer  9208  transmits the accession numbers to process controller  26 , which sends them to LIMS  24 . In this manner, LIMS  24  maintains a record of each sample and its location in each source well container  2  and in each primary master well container  6 . LIMS  24  and process controller  26  correlate the accession number of each primary master well container  6  with the identity of each sample it contains, the strain for which each sample is to be tested, the designated genetic sequence or sequences that identify or indicate that strain, the probes and primer sets necessary to test for those designated genetic sequences and the results of the testing.  
      The tray of primary master well containers is moved by the transportation apparatus to the Isolation/Purification Station  94 . In this station, the genomic nucleic acid will be isolated and purified using a separation method such as magnetic or paramagnetic particles. Purified genomic nucleic acid, substantially free of protein or chemical contamination is obtained by adding a sufficient amount of magnetic particles to each of the well containers that bind to a predefined quantity of nucleic acid. The term “magnetic” in the present specification means both magnetic and paramagnetic. The magnetic particles can range from 0.1 micron in mean diameter to 100 microns in mean diameter. The magnetic particles can be functionalized as shown by Hawkins, U.S. Pat. No. 5,705,628 at col. 3 (hereinafter &#39;628 patent hereby incorporated by reference).  
      In the preferred embodiment, the magnetic particles are purchased from Promega Corporation, a measured amount of magnetically responsive particles are added  133  to the lysate mixture with or without the presence of a chaotropic salt  135 . In the preferred embodiment, 13 μl amounts of 1 micron silica magnetic particles with chaotrope 113 μl (Promega Corporation, Madison, Wis.) are added to each well of the microwell container. The fixed volume of particles becomes saturated with nucleic acid if there is enough nucleic acid in the lysate. It has been observed that the resulting nucleic acid concentration between samples is very consistent if there is an excess nucleic acid is present in the lysate. In a 50 μl pathlength read by the Genios (Tecan, Research Triangle Park, N.C.) a standard A 260  is 0.2 OD units. A standard concentration range of 0.1 to 0.3 O.D. units is disassociated from the magnetic particles to yield purified genomic nucleic acid.  
      Table 1 shows that with increasing amounts of magnetic particles, the nucleic acid concentration also increases.  
                       TABLE 1                               Bead Volume               per       Average   Stdev   150 μl of lysate                                            0.7974   0.0072   27       0.8750   0.040   35       1.2328   0.026   50       1.7900   0.022   75                  
 
      While the nucleic acid concentration is consistent between samples treated with the same protocol, several factors may increase or decrease the resulting standard concentration of genomic nucleic acid. These factors include: the starting amount of nucleic acid in each lysate preparation, the binding reagent, the number of purification washes, and the solution that is used to elute the nucleic acid. The preferred binding solution for the magnetic particles obtained from Promega (Madison, Wis.) is a chaotropic salt, such as guadinium isothiocyanate. Alternatively, other binding reagents, such as 20% polyethylene glycol (PEG) 8000, 0.02% sodium azide and 2.5M sodium chloride may be used to nonspecifically bind the genomic nucleic acid to the surface chemistry of the functionalized magnetic particles. If functionalized magnetic particles are used, the preferred binding solution is PEG. The PEG or chaotropic guadinium isothiocyanate allows for the disruption of hydrogen binding of water, which causes binding of the nucleic acid to the particles. The preferred washing procedure to remove contaminants includes two chaotrope washes, after the initial chaotrope binding step, followed by four 95% ethanol washes. Aqueous solutions, or the like, are the best elution solutions. These solutions include water, saline sodium citrate (SSC) and Tris Borate EDTA (ie. 1×TBE).  
      The amount of DNA isolated from the swabs and blood is less than the DNA yield recovered from tissue. The tissue lysate has enough DNA content to saturate the binding ability of the fixed volume of beads. However, the swab and blood lysate does not have enough DNA to saturate the binding ability of the fixed amount of beads. This is evidence by the CT (cycle threshold) values for the housekeeping probe. The housekeeping (cjun) CT values for tissue isolations are approximately 26 whereas the approximate CT for housekeeping (cjun) for the blood isolations are approximately 35. This nine cycle difference represents approximately a 512 (2ˆ9) fold difference in the amount DNA present. This non-saturated DNA yield does not present a problem for results because the housekeeping probe normalizes the results. For each sample, the CT values for the wells containing the housekeeping probe, cjun, are averaged (CT cjun ). The RCN (RCN 1  and RCN 2 ) values are calculated by comparing the test probe (i.e. Cre or MN1TEL) signal to the housekeeping gene signal average for each of the two test probe wells (CT 1  and CT 2 ), the following equation is applied: 
 
 RCN   1 =2 −(CT     1     −CT     cjun     )  
 
 RCN   2 =2 −(CT     2     −CT     cjun     )  
 
      The preferred device for performing the above functions of the Isolation/Purification Station  94  is a liquid handler  9402  identical in general construction to the liquid handler  9220  identified above for use as the Lysing Station  92  and the Accessioning Station  93  that has been configured to automatically transfer the various reagents and other liquids as well as the magnetic particles in the manner described below.  
       FIG. 16  illustrates a preferred embodiment of the liquid handler  9402 . Handler  9402  comprises a frame  9404  on which is mounted a deck  9406 , which is surmounted by material handler  9408 , which supports and positions pipettes  9410  and is coupled to and controlled by computer  9412 , which is in turn coupled to process controller  26  to communicate information to and from LIMS  24 . Liquid handler  9402  includes a syringe pump  9414  that is coupled to and driven by computer  9412  to dispense magnetic particles via a 16×24 array of 384 pipettes  9410  simultaneously into all 384 wells of the primary master well container  6  under the command of computer  9412 . Liquid handler  9402  also includes a second syringe pump  9416  that is configured to dispense a binding buffer into wells of the primary master well container  6  under computer control. The liquid handler also includes a magnet  9418  mounted in deck  9406  as well as a conveyor  9420  that is coupled to and controlled by computer  9412  to move the primary master well container  6  in tray  9206  back and forth between a first position  9422  in which the container is within the magnetic field and a second position  9424  in which the container is outside the magnetic field.  
      Before the functions of the Isolation and Purification Station  94  can be performed, the operator must first move the primary master well container  6  from Accessioning Station  93  to deck  9406  of liquid handler  9402  and place it in a predetermined location on the deck. Once the operator has placed the primary master well container  6 , the operator starts an isolation/purification program running on computer  9412 . This program drives the operations of liquid handler  9402  causing it to dispense magnetic particles  133  into all the wells of the primary master well container  6  containing lysed samples. Computer  9412  signals syringe pump  9414  to dispense the particles using pipettes  9410  into the primary master well container  6  when container  6  is in position  9424 , away from the magnetic field created by magnet  9418 .  
      Once the particles have been added, computer  9412  then directs the pipettes  9410  to add a chaotropic salt such as guadinium isothiocyanate to each of the wells to bind the genomic nucleic acid to the magnetic particles at  135 . Once the chaotropic salt has been added, computer  9412  then mixes the contents of the wells by signaling the pipettes  9410  to alternately aspirate and redispense the material in each of the wells. This aspiration/redispensing process is preferably repeated three or four times to mix the contents in each well.  
      Once the contents of the wells have been mixed, computer  9412  pauses for two minutes to permit the particles, binding reagent, and raw biological material in the wells to incubate at room temperature in position  9424 . When the two minutes have passed, computer  9412  commands the conveyor  9420  to move tray  9206  from position  9424  to position  9422 , directly above magnet  9418  at  137 . In this position the magnet draws the magnetic particles in each of the wells downward to the bottom of the wells of the primary master well container  6 . Computer  9412  keeps tray  9206  and the primary master well container  6  over the magnet and within the magnetic field for 2-6 minutes, or until substantially all the magnetic particles are drawn to the bottom of each well and form a small pellet.  
      The particles drawn to the bottom of each well have genomic nucleic acid attached to their outer surface—genomic nucleic acid that the particles hold until an elution solution is placed in each well to release the genomic nucleic acid from the particles. With the particles at the bottom of each well and the wells located within the magnetic field, computer  9412  directs the pipettes to aspirate the supernatant  139 .  
      Once the supernatant is removed, computer  9412  signals the conveyor to move the primary master well container  6  on tray  9206  to the nonmagnetic position  9424 . The foregoing process of adding chaotropic salt, mixing the combination, pausing, drawing the magnetic particles down and aspirating the supernatant is repeated two more times.  
      Computer  9412  then directs the pipettes to introduce a wash solution (for example 70% ethanol when functionalized beads are used, or 95% ethanol (4×) when silica beads are used) to resuspend the particles  141 . Computer  9412  again mixes the contents of the wells by signaling the pipettes to alternately aspirate and redispense the material in each of the wells. With the wash buffer and particles thoroughly mixed, computer  9412  again moves tray  9206  and the primary master well container  6  back over magnet  9420  in position  9422   143  and draws the magnetic particles back to the bottom of the wells. This wash process  141 , 143 , 145  is repeated three times to thoroughly cleanse the magnetic particles, and dilute and remove all supernatant.  
      Once the particles are thoroughly washed, computer  9412  permits the magnetic particles in each well to air dry  147 . In the preferred embodiment, shown in  FIG. 17 , the operator moves the primary master well container  6  to a dryer  9426  (an “Ultravap” dryer by Porvair Sciences, UK) having 384 tubules disposed in a 16×24 array  9428  that are configured to be simultaneously inserted into each of the wells of the primary master well container  6  and to supply warm, dry air thereto. In an alternative method, computer  9412  causes material handler  9408  to direct compressed dry nitrogen gas into each well of the primary master well container  6 , drying the particles out in place while the container is in the magnetic field. Alternatively the samples can be permitted to air dry. Once the particles are completely dry, the primary master well container  6  can be subsequently moved away from the field of magnet  149 .  
      Once the particles are almost dry, the operator returns the primary master well container  6  to the liquid handler  9402  and directs the computer  9412  to command the pipettes  9410  to fill the wells with an elution solution  151  and resuspend the particles. This elution solution is formulated to elute the bound genomic nucleic acid from the particles. An example of one such elution solution is 0.01M Tris (pH 7.4), sodium saline citrate (SSC), dimethyl sulfoxide (DMSO), sucrose (20%), 1×TBE, or formamide (100%). In the preferred embodiment, the elution solution is nuclease-free water. Nuclease free water is selected to minimize contamination and produce a standard concentration of purified genomic nucleic acid. In the preferred embodiment, the elution solution temperature is 22° C. A preferred yield is about 20 ng/μL of genomic nucleic acid is obtained.  
      After resuspending the genomic nucleic acid in a solution for a predetermined period of time, computer  9412  again moves tray  9206  with the primary master well container  6  via conveyor  9420  to position  9422  over magnet  9418   155 . The magnet, in turn, draws the magnetic particles down to the bottom of each well. This leaves the genomic nucleic acid mixed and suspended in the elution solution. Computer  9412  then directs the pipettes to aspirate a small amount (50 μl) of purified genomic nucleic acid and to transfer  159  the small amount from each well into a corresponding well of a clean optical 384-well container that is also mounted on deck  9406 . The operator scans  161  a barcode accession number on the optical container and computer  9412  transfers the scanned accession number to process controller  26 , which then transfers it to LIMS  24 . The operator takes this optical container to a UV spectrometer (Genios, by Tecan of Raleigh-Durham, N.C.), and directs the UV spectrometer to optically scan the optical container, by making an A 260  measurement  163 . This measurement is electronically transferred  112  to LIMS  24  over a data communications link.  
      If another fully automated system is desired, the magnetic separator can be automated and rise from the bottom of the workstation and make contact with bottoms of all primary well containers simultaneously.  
      In the preferred embodiment for the biological sample, the genomic nucleic acid is not sonicated after separation from the cellular debris. The genomic nucleic acid includes at least a portion of intact nucleic acid. Unsonicated nucleic acid is recovered in the condition it is found in the lysate. Thus, if the genomic nucleic acid is intact in the lysate, it is intact (i.e., unfragmented) as attached to the particles. The sample contains at least a portion of intact genomic nucleic acid.  
      In certain types of samples, such as embryos, the genomic nucleic acid is substantially intact. In one embodiment, the genomic nucleic acid can be sonicated before or after separation with the magnetic particles. When the biological tissue is embryonic sonication is preferred. Sonication can be done by any conventional means such as a fixed horn instrument or plate sonicator. In the one embodiment, the genomic nucleic acid is sonicated for five seconds to produce nucleic acid fragments. Although there is a wide range of fragments from about 100 base pairs to up to 20 kilobases, the average size of the fragment is around about 500 base pairs.  
      The primary master well container  6  is transported to the deck of the Screening Station  95  ( FIG. 18 ) where its bar code is scanned  173 . The operator places the container on a magnet, drawing all the magnetic particles to the bottom of the wells. The supernatant contains the purified genomic nucleic acid. LIMS  24  generates a worklist containing barcodes that list the primer/probe combinations that need to be loaded onto the deck of the machine. The primer-probe combinations are contained in barcoded tubes. An operator loads the barcoded tubes randomly into a probe box. The operator then scans the barcodes on the tubes using a Matrix scanner coupled to LIMS  24 . The primer set and probe combinations in the tubes are then loaded into an ABI 384 PCR plate (Applied Biosystems, Forest City, Calif.). The genomic nucleic acid sample from each well of the primary master well container  6  is added to a corresponding well of the ABI PCR plate that contains the primer-probe combination or combinations appropriate to discern the relevant genotype  187 . The ABI plate is then sealed with sealing tape and taken to the Detection Station  96  and placed in an ABI 7900. In the preferred embodiment the ABI 7900 cycles the ABI PCR plate 40 times between temperatures specified by the manufacturer. The operator can vary the number of cycles and the temperatures as desired to increase the signal provided by the samples.  
       FIG. 18  shows a preferred device for performing the Screening Station  95  functions. It comprises a liquid handler  9502  such as Genesis Tecan (Raleigh Durham, N.C.) or Multimeck Beckman (Indianapolis, Ind.). It includes a frame  9504 , on which a deck  9506  is mounted to provide a horizontal working surface for first tray  9206  and second tray  9206 . The first and second trays (as described above) can support and position nine primary master well containers  6 .  
      Liquid handler  9502  also includes a material handler  9508  that is fixed to frame  9504  and extends upward and across the top surface of deck  9506 . A computer  9510  is coupled to material handler  9508  to direct the movement and operation of pipettes  9512 . Pipettes  9512  are fluidly coupled to a syringe pump  9514 .  
      Probe block  9516  is disposed on the surface of deck  9506  and contains several tubes (not shown) each tube containing one or more combined primer sets and probes. The operator bar-codes each tube and enters the data indicative of the tube contents (the particular primer or probe in each tube, its volume and concentration) into LIMS  24 , which stores the data associated with the bar code on the tube for later reference  173 .  
      The operator places the primary master well containers  6  on deck  9506 , scans the bar code accession number of the primary master well container  6 , and signals computer  9510  to start transferring genomic nucleic acid, probes and primer sets.  
      Based upon the information provided by the remote user  1 , including the samples, the strains for which the samples are to be tested, and the designated genetic sequences indicated by the strains, as well as the probes and primer sets necessary to detect those designated genetic sequences, as well as the location of each sample in the ABI PCR plate, LIMS  24  calculates a worklist that identifies for the operator which (and how many) tubes containing which probes and which primer sets must be placed in the probe block  9516  to test the samples in the primary master well container  6 .  
      The operator first prints out this worklist, using it as a guide to identify and select particular tubes containing the proper probes and primers. The operator takes these tubes out of storage, places them in the probe block  9516  and places the probe block  9516  on the Matrix scanner.  
      The Matrix scanner is coupled to LIMS  24 , and is configured to scan the bar codes on each tube through holes in the bottom of the probe block. The scanner passes this information to LIMS, to which it is coupled, which in turn compares the bar codes of the scanned tubes with the bar codes of the probes identified on the worklist. Only if the operator has loaded the probe block with the appropriate type and number of probes and primer sets will LIMS  24  permit the operator to proceed. In this manner, LIMS is configured to verify that the operator has inserted the appropriate and necessary tubes of probes and primer sets into the probe block.  
      Once LIMS  24  has verified that the proper tubes of probes and primer sets have been inserted into the probe block, it is configured to indicate to the operator that the probe block is acceptable and that the process steps at Screening Station  95  can begin.  
      The steps of preparing tubes of probes and primer sets, entering them into LIMS, preparing a worklist, filling a probe block and verifying the probe block, all happen prior to the time the operator takes the primary master well container  6  with its 384 wells to the deck  9506  of liquid handler  9502  and places it in position on deck  9506 .  
      The operator places the primary master well container  6  in position on first tray  9206  located on deck  9506  of liquid handler  9502 . The operator electronically scans the container with an electronic scanner  9518  coupled to computer  9510  which, in turn, is coupled to process controller  26 . As described above, the scanner may be any of several types of electronic scanner but is preferably a bar code scanner.  
      If there are several primary master well containers  6 , they are preferably carried from the liquid handler of the Isolation/Purification Station  94  to the liquid handler of the Screening Station  95  in tray  9206 , which can accommodate nine separate primary master well containers  6 .  
      The operator also places a secondary master well container  27  (preferably an ABI 384 PCR plate) in a predetermined location on the second tray  9206  located on deck  9506  adjacent to the first tray  9206 . The operator electronically scans the secondary master well container  27  with the electronic scanner  9518  and stores the location and identity of the secondary master well container  27  in process controller  26  which transmits the data to LIMS  24 .  
      If there are several primary master well containers  6  that must be transferred to secondary master well containers  27 , the corresponding secondary master well containers  27  may also be taken to liquid handler  9502  in trays  9206 , rather than the operator carrying each secondary master well container  27  to second tray  9206  individually.  
      Once the operator places at least one primary master well container  6  in first tray  9506  and at least one secondary master well container  27  in second tray  9506 , the operator signals computer  9510  to begin combining the probes, primer sets, and genomic nucleic acid extracted from the samples.  
      Generally speaking, computer  9510  commands material handler  9508  to extract probes and primer sets from tubes in probe box  9516  and deposit them in each secondary master well container  27  in second tray  9206 . Computer  9510  then commands material handler  9508  to extract the genomic nucleic acid from the wells of each primary master well container  6  in first tray  9206  and deposit the samples in wells in a corresponding secondary master well container  27 . When the pipettes  9512  deposit the genomic nucleic acid samples, the probes, and the primer sets in wells in the secondary master well containers  27 , computer  9510  commands material handler  9508  and pipettes  9512  to mix the samples using the aspiration/redispensing methods discussed above.  
      The secondary master well containers  27  receive a number of aliquots of biological sample in multiple wells of the secondary master well container. In one embodiment, an aliquot of the biological sample of the strain is dispensed into at least four wells of the secondary master well container  27 . To at least two of the four wells at least one probe and primer set (e.g. SEQ ID NO. 23, 24 &amp; 25) corresponding to at least one designated genetic sequence is added. A probe (SEQ ID NO. 21) and primer set (SEQ ID NO. 19 &amp; 20) correspond to a reference sequence (SEQ ID NO. 18) is added to the third and fourth well. Thus, for example, if the genotype screening includes four designated genetic sequences, then four wells of the secondary master well containers  27  receive an aliquot of the biological sample and the corresponding probes and primer sets for each designated genetic sequence. Additionally, four wells receive an aliquot of the biological sample and the corresponding four probe and primer sets. This second set of wells is referred to as the replicants. The function of the replicants is quality control. Additionally, two additional wells receive aliquots of the biological sample and the housekeeping or screening reference probe/primer set.  
      In a simpler embodiment, the validity of the screening data can be evaluated by dispensing an aliquot of a biological sample of the strain designated by the remote user into at least two wells of a microwell container. In one well at least one probe and primer set is added corresponding to the at least one designated genetic sequence and to the other well at least one probe and primer is added corresponding to the reference sequence (SEQ ID NO. 18). The biological sample is screened and the probe signal values are compared between the probe for the designated genetic sequence and the probe for the referenced sequence.  
      In other embodiments, multiple probe and primer sets can be multiplexed into a single well. Furthermore, the detection of SNPs involve adding two probes to a well.  
      Between one and five microliters of nucleic acid and four and fifteen microliters of probes and primer sets are preferred to insure proper mixing of the samples and proper polymerization in the PCR process of the Detection Station  96  that follows.  
      Once the wells in the secondary master well containers  27  are filled with the appropriate purified genomic nucleic acid samples, primer sets and probes, and these materials are mixed, computer  9510  signals the operator that the screening process is complete. The plate is then sealed with optical sealing tape. The operator then moves the secondary master well containers  27  to Detection Station  96  for further processing.  
      In the preferred embodiment, the central component of Detection Station  96  is the ABI 7900. The secondary master well containers  27  are placed inside the ABI 7900, where they are thermocycled  189  40 times and exposed to an excitatory energy source to produce a quantifiable signal  195  from the signal molecule. More particularly, the Detection Station  96  scans the secondary master well container&#39;s  27  barcode and reports it  196  to LIMS  24 .  
       FIG. 19  illustrates a preferred device for performing the functions of Detection Station  96 . It includes a PCR instrument  9602  (here shown as an ABI 7900), a material handler  9604  (here shown as a ZYmark arm), a computer  9606 , and an electronic scanner  9608  (here shown as a barcode scanner).  
      Computer  9606  is coupled to PCR instrument  9602 , material handler  9604 , and process controller  26 . It communicates with PCR instrument  9602  to control the insertion and removal of secondary master well containers  27  from PCR  9602  by handler  9604 . Computer  9606  is also coupled to PCR instrument  9602  to process test results from the test performed by PCR instrument  9602  and to transmit those test results to process controller  26  and then to LIMS  24 .  
      Scanner  9608  is coupled to handler  9604  to scan the accession numbers on the secondary master well containers  27 , and to transmit those accession numbers to LIMS  24 .  
      Material handler  9604  includes an arm  9610  that is commanded by computer  9606  to move between three positions: an incoming material hopper  9612 , and outgoing material hopper  9614 , and loading/unloading position  9616 . Handler  9604  moves between these positions under the control of computer  9606 , which commands this movement.  
      The operator first loads incoming material hopper  9612  with one or more secondary master well containers  27 . The operator then operates the computer terminal  9618  of computer  9606 , commanding computer  9606  to load and test the secondary master well containers  27 . In response, computer  9606  commands arm  9610  to move to the incoming material hopper  9612 , grasp the topmost secondary master well container  27 , and to carry that container to the loading/unloading position  9616 . Computer  9606  also commands PCR instrument  9602  to extend a tray (not shown) from an opening  9618  in the side of the ABI 7900, and commands arm  9610  to place the secondary master well container  27  on that tray. Scanner  9608  is configured to scan the barcode accession number on the secondary master well container  27 , thereby making an electronic record of the secondary master well container  27  that is being tested. Scanner  9608  transmits this accession number to computer  9606 , which later correlates the accession number with the test results provided by ABI 7900.  
      Once the secondary master well container  27  is placed in the tray, computer  9606  commands PCR instrument  9602  to retract the tray, and to begin testing the material in the secondary master well container  27 , which is now inside PCR instrument  9602 . PCR instrument  9602  signals computer  9606  when testing is complete. PCR instrument  9602  also transmits the test results to computer  9606 . Computer  9606 , in turn, commands PCR instrument  9602  to eject the secondary master well container  27  that has just been tested, moving it back to loading/unloading position  9616 . Once the secondary master well container  27  is in this position, computer  9606  commands material handler  9604  to move arm  9610  back to the loading/unloading position  9616  and to retrieve the secondary master well container  27  that has just been tested. Computer  9606  commands arm  9610  to move the just-tested secondary master well container  27  to outgoing material hopper  9614 , where it is deposited, awaiting later removal by the operator of Detection Station  96 .  
      Now referring to  FIG. 9 , LIMS  24  now prepares the outcome report  249 . Several calculations are performed before they are posted to the outcome report  249 . In the preferred embodiment, such calculations include the evaluation of all replicates per sample. Calculating the relationship between the experimental quantified signal and the quantified signals of designated control may elucidate the copy number, zygosity or mosaic nature of the sample. The ratio for homozygous individuals should be twice the ratio of heterozygous individuals.  
      A reference sequence (SEQ ID NO. 18) and respective primer set and probe (SEQ ID NO. 19-21) is used to normalize the signal of every other probe used for that sample. The resulting value, called an RCN, is a comparison of the signal of the test probe (i.e. probes for portion of the designated genetic sequences) to the reference sequence. This control serves an additional purpose which is to evaluate the consistency of the nucleic purification system. This control will produce a magnitude of fluorescence directly proportional to the amount of starting nucleic acid, so nucleic acid concentrations can be compared. More specifically, the probe value corresponds to the designated genetic sequence is compared to the probe value of the replicant. Similarly, each value is compared to the probe value for the reference sequence to evaluate the validity of the data obtained.  
      For each sample, the CT values for the two wells containing the housekeeping gene, cjun, are averaged (CT cjun ). The RCN values are calculated by comparing the test probe (i.e. Neo or Cre) signal to the housekeeping gene signals or each of the two test probe wells (T 1  and T 2 ), the following equation is applied:  
               TABLE 2                          Example of RCN Calculation       RCN 1  = 2 −(CT     1     −CT     cjun     )         RCN 2  = 2 −(CT     2     −CT     cjun     )                                                               Average           Well   Sample Name   Detector   Task   CT   c-jun   RCN               C1   Neomycin KO 1   c-jun   Unknown   25.37   25.27           D1   Neomycin KO 1   c-jun   Unknown   25.17       E1   Neomycin KO 1   Neo A   Unknown   33.27       0.00       F1   Neomycin KO 1   Neo A   Unknown   34.24       0.00                  
 
      Now referring to  FIG. 9 , the sample outcome report  249  may include account registration  250 , well plate container  2  barcode number(s) (i.e. accession numbers)  252 , control sample locations  252  and genetic characterization of the designated control  252 . Additionally, the outcome report  249  may include well location  254 , sample identification  256 , nucleic acid concentration  260 , signal quantification  266 , qualitative results  268 , zygosity/copy number  270 , quantitative analysis via comparison to designated control signal strengths allowing for copy number estimation, zygosity or mosaic nature  270 . The outcome report  249  may also include a picture file (email) or pictorial representations of results  272  as shown in  FIG. 10 . Additionally, information gathered at the request of the remote user  1  from optimization and sequence confirmation quality control data and error messages may be included in the outcome report  249 . The remote user  1  may choose to have this file electronically sent or choose to be electronically notified. Additionally, remote user  1  has the option to have a hard copy sent via the postal service or facsimile.  
      Once the LIMS  24  has compiled all the data for the outcome report  249 , the outcome report will be sent  7  to the remote user  1 . In the preferred embodiment, LIMS  24  will send the report via a remote link  7  to either the remote user  1  or the order manager  22 , which can post the results on the web site  16  or via an electronic link  7 . The LIMS  24  will keep results available for six months and then the results will be recorded onto a long-term storage disk and archived.  
      The following examples are provided by way of examples and are not intended to limit the scope of the invention.  
     8. EXAMPLES  
     Example 1  
     Swab Sample Collection Method  
      MasterAmp Nylon Buccal Swabs (MB030BR Epincentre, Madison, Wis.), Microbrushes (MG-400, Sullivan Schein Melville, N.Y.) and Proxabrush conical brushes (618PNE GUM, Chicago, Ill.) are used to collect DNA samples from the oral or nasal cavity as well as the anal region of mutant and wild type mice. The swabs tips were removed and placed in individual wells of a VWR-DYNBL deep 96 well plate. One hundred fifty microliters of SV Lysis reagent (Promega Corporation Z305X) is added to each well containing a sample. The swabs are then incubated at room temperature for ten minutes. The well plate is then placed back on the deck of the Tecan Genesis Workstation. The liquid handler aspirates 100 μl of each sample and dispenses it in to a 384 well-plate primary master well container. Once all of the samples are transferred, the primary master well container is moved to the deck of the Isolation station Purification Station  94 .  
      Fifty microliters of SV Lysis reagent (Z305X Promega Corporation, Madison, Wis.) are added to each sample. Next, 13 μl of magnetic particles (Promega Corporation A220X) are added and the well components are mixed. The well plate is then moved into a magnetic field where the magnetic particles (Promega Corporation #A220X) are drawn to the bottom of each well. The supernatant is then aspirated and discarded. The well plate is moved out of the magnetic field and 113 μl of SV Lysis reagent is added to each well and mixed. The microwell container is then moved into the magnetic field and the supernatant was drawn off and discarded. Next, the sample is washed two times in 125 μl of 95% ethanol as described above. After the second ethanol wash, the microwell container is placed on a 384 tip dryer for 11 minutes. Then the microwell container is moved back to the deck of the Isolation/Purification  94  station and 155 μl of Ambion&#39;s (Houston, Tex.) nuclease free water (catalog #B9934) is added to each well at room temperature. The microwell container is then moved into the magnetic field and 50 μl of DNA is transferred to a 384 well optical storage plate (Fisher Scientific, #08-772136) for optical density analysis.  
      The primary master wellplate with the isolated DNA is moved to the deck of a Tecan Freedom Workstation. The TaqMan Universal Master Mix, real time-PCR primer set/probe (for the designated genetic sequence) mixture and Ambion water are added to the microwell container. The final PCR mixture is made of 1× TaqMan Universal Master Mix (catalog # 4326708), 1× real time PCR primer mix (Applied Biosystems Assays-by-Design(SM) Service 4331348) and 25% isolated DNA. The Tecan Genesis adds the reagents together in the ABI 7900 384 Well Optical Plate. The plate is then sealed with optical sealing tape (#4311971, Applied Biosystems). The samples are then placed in an Applied Biosystems SDS HT7900. A standard real time PCR protocol is followed by heating the samples to 50° C. for two minutes, holding the samples at 95° C. for ten minutes, followed by thermally cycling the sample 50 times between 95° C. for 15 seconds and at 60° C. for one minute.  
      The results are shown in Tables 3 and 4.  
                           TABLE 3                               Designated                   Genetic       Well   Sample Name   Sequence   CT                                                C4   Blue GUM   Cjun   33.86       D4   Blue GUM   Cjun   34.23       E4   Blue GUM   Neomycin   30.22       F4   Blue GUM   Neomycin   30.08       C1   GUM 1   Cjun   32.56       D1   GUM 1   Cjun   32.22       E1   GUM 1   Neomycin   28.03       F1   GUM 1   Neomycin   28.01       C3   GUM 2   Cjun   33.2       D3   GUM 2   Cjun   33.23       E3   GUM 2   Neomycin   28.95       F3   GUM 2   Neomycin   29.08       C6   MasterAmp 1   Cjun   31.77       D6   MasterAmp 1   Cjun   31.7       E6   MasterAmp 1   Neomycin   27.45       F6   MasterAmp 1   Neomycin   27.56       G6   MasterAmp 2   Cjun   30.6       H6   MasterAmp 2   Cjun   30.68       A7   MasterAmp 2   Neomycin   26.72       B7   MasterAmp 2   Neomycin   26.67       G1   Micro Green 1   Cjun   31.42       H1   Micro Green 1   Cjun   31.76       A2   Micro Green 1   Neomycin   26.09       B2   Micro Green 1   Neomycin   26.15       G2   Micro Green 2   Cjun   33.31       H2   Micro Green 2   Cjun   33.74       A3   Micro Green 2   Neomycin   29.1       B3   Micro Green 2   Neomycin   29.2       G3   Micro Green 3   Cjun   32.91       H3   Micro Green 3   Cjun   33.12       A4   Micro Green 3   Neomycin   28.73       B4   Micro Green 3   Neomycin   29.03       C5   Micro Green 4   Cjun   35.25       D5   Micro Green 4   Cjun   35.1       E5   Micro Green 4   Neomycin   31.23       F5   Micro Green 4   Neomycin   30.95       G5   Micro Green 5   Cjun   34.39       H5   Micro Green 5   Cjun   34.84       A6   Micro Green 5   Neomycin   30.49       B6   Micro Green 5   Neomycin   30.62       G4   Micro Yellow   Cjun   32.8       H4   Micro Yellow   Cjun   32.88       A5   Micro Yellow   Neomycin   29.12       B5   Micro Yellow   Neomycin   28.9       C2   Whatman   Cjun   34.05       D2   Whatman   Cjun   34.04       E2   Whatman   Neomycin   29.21       F2   Whatman   Neomycin   29.4       A1   Water   Cjun   Undetermined       B1   Water   Cjun   Undetermined                  
 
     
       
         
           
               
               
               
               
               
             
               
                   
                 TABLE 4 
               
             
            
               
                   
                   
               
               
                   
                   
               
               
                   
                 Rep1 
                 Rep 2 
                 CJUN 
                 NEO 
               
            
           
           
               
               
               
               
               
            
               
                   
                 RCN 
                   
                 CT 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Blue GUM 
                 14.17 
                 15.62 
                 34.0 
                 30.2 
               
               
                   
                 GUM 1 
                 20.53 
                 20.82 
                 32.4 
                 28.0 
               
               
                   
                 GUM 2 
                 19.23 
                 17.57 
                 33.2 
                 29.0 
               
               
                   
                 MasterAmp 1 
                 19.49 
                 18.06 
                 31.7 
                 27.5 
               
               
                   
                 MasterAmp 2 
                 15.14 
                 15.67 
                 30.6 
                 26.7 
               
               
                   
                 Micro Green 1 
                 45.25 
                 43.41 
                 31.6 
                 26.1 
               
               
                   
                 Micro Green 2 
                 21.48 
                 20.04 
                 33.5 
                 29.2 
               
               
                   
                 Micro Green 3 
                 19.49 
                 15.83 
                 33.0 
                 28.9 
               
               
                   
                 Micro Green 4 
                 15.40 
                 18.70 
                 35.2 
                 31.1 
               
               
                   
                 Micro Green 5 
                 17.45 
                 15.94 
                 34.6 
                 30.6 
               
               
                   
                 Micro 
                 13.18 
                 15.35 
                 32.8 
                 29.0 
               
               
                   
                 Yellow 
               
               
                   
                 Whatman 
                 28.54 
                 25.02 
                 34.0 
                 29.3 
               
               
                   
                   
               
            
           
         
       
     
     Example 2  
     Blood Sample Collection Method  
      Mouse tails are nicked with a razor blade and the resulting blood droplets are blotted on to filter paper (V&amp;P Scientific Lint Free Blotting Media (114 mm long, 74 mm wide) #VP540D). The samples are placed in individual wells of a Nunc 96-well plate (Fisher Scientific 12-565-368). The well locations are labeled and the plates are transported shipped to the screening laboratory  20 .  
      The remote user  1  provides the genetic line identification  84 . The genetic line in this example has been previously associated by the remote user  1  with the designated genetic sequence for MnlTel (SEQ ID NO. 38), CRE (SEQ ID NO. 22) and MHV (SEQ ID NO. 34).  
      The number of samples are counted and lysis reagent is made (2.5 μl of proteinase K (VWR EM-24568-3) and 147.5 μl of Nuclei Lysing Solution (Promega Corporation, Madison Wis., A7943) per sample. The solution is gently mixed and poured into a 25 ml trough or reservoir and placed on the deck of a Tecan Genesis Workstation (Research Triangle Park, N.C.). The liquid handler dispenses 150 μl of the solution into each sample well. The well plate is then placed in a 55° C. oven for three hours.  
      The well plate is then placed back on the deck of the Tecan Genesis Workstation. The liquid handler aspirates 50 μl of each sample and dispenses it in to a 384 primary master well container (Fisher Scientific #NC9134044). Once all of the samples are transferred, the primary master well container is moved to the deck of the Isolation Station Purification Station  94 .  
      One-hundred and twelve microliters of SV Lysis reagent (Promega Corporation, # Z305X) are added to each sample. Next, 13 μl of magnetic particles (Promega Corporation # A220X) are added and the well components are mixed. The well plate is then moved into the magnetic field of a magnet where the magnetic particles are drawn to the bottom of each well. The supernatant is then aspirated and discarded. The well plate is moved out of the magnetic field and 95 μl of SV Lysis reagent is added to each well and mixed. The well plate is then moved into the magnetic field and the supernatant is drawn off and discarded. This washing process is repeated two additional times. Next, the samples are washed four times in 130 μl of 95% ethanol as described above. After the last ethanol wash, the well plate is placed on a 384 tip dryer for 11 minutes. Then the well plate is moved back to the deck of the Isolation Station and 155 μl of Ambion&#39;s (Houston, Tex.) nuclease free water (catalog #B9934) is added to each well. The elution solution is heated to 95°. The plate is then moved into the magnetic field and 50 μl of DNA elution is transferred to a 384 well optical storage plate (Fisher Scientific, #08-772136) for optical density analysis.  
      An A 260  reading of the storage plate read is performed with a Tecan Genios Spectrometer. This reading shows nucleic acid is present at the desired concentration of 0.2 O.D. units, but, a range of 0.1 to 0.5 O.D. units is acceptable.  
      The plate with the isolated DNA is moved to the deck of a Tecan Freedom Workstation; TaqMan Universal Master Mix, real time PCR primer mixture and Ambion water are placed on the deck as well. The final PCR mixture is made of 1× TaqMan Universal Master Mix (catalog # 4326708), 1× real time PCR primer mix for a designated genetic sequence (Applied Biosystems Assays-by-Design(SM) Service 4331348) and 25% isolated genomic DNA.  
      In this example, the primer set as set out in SEQ ID NO. 23 and 24 and probe as set out in SEQ ID NO. 25 correspond to the designated genetic sequence CRE (SEQ ID NO. 22). Additionally, the primer set as set out in SEQ ID NO. 35 and 36 and probe as set out in SEQ ID NO. 37 correspond to the designated genetic sequence MnlTel (SEQ ID NO. 38). Additionally, the primer set as set out in SEQ ID NO. 35 and 36 and probe set out as set in SEQ ID NO. 37 corresponds to the designated genetic sequence MHV (SEQ ID NO. 34).  
      The Tecan Genesis adds the reagents together in the ABI 7900 384 Well Optical Plate (Foster City, Calif.) catalog #4309849). The 384 well plate is then sealed with optical sealing tape (ABI, #4311971).  
      The samples are then placed in an Applied Biosystems SDS HT7900 (Foster City, Calif.). A standard real time PCR protocol is followed by heating the samples to 50° C. for two minutes then incubated at 95° C. for 10 minutes, followed by thermally cycling the samples 40 times between 95° C. for 15 seconds and 60° C. for one minute.  
               TABLE 5                          Blood Samples Taken from Double KO mice       Whatman Filter Paper used to capture samples                                                 Designated                       Sample   Genetic       Std. Dev.           Well   Name   Sequence   CT   CT                                                     A1   WATER   Cjun   Undetermined                                             A2   Blood 2   Cjun   35.31   0.587           A3   Blood 3   MN1TEL   33.51   0.061           A4   Blood 4   CRE   34.72   0.27           A5   Blood 6   Cjun   35.78   0.175           A6   Blood 7   MN1TEL   33.24   0.325                                         A7   Blood 8   CRE   Undetermined                                             A8   Blood 10   Cjun   35.44   0.023           A9   Blood 11   MN1TEL   35.25   0.004           A10   AF 2   Cjun   37.25   0.786           A11   AF 4   Cjun   35.17   0.165                                         B1   WATER   Cjun   Undetermined                                             B2   Blood 2   Cjun   34.48   0.587           B3   Blood 3   MN1TEL   33.42   0.061           B4   Blood 4   CRE   34.34   0.27           B5   Blood 6   Cjun   36.03   0.175           B6   Blood 7   MN1TEL   33.7   0.325                                         B7   Blood 8   CRE   Undetermined                                             B8   Blood 10   Cjun   35.47   0.023           B9   Blood 11   MN1TEL   35.25   0.004           B10   AF 2   Cjun   36.14   0.786           B11   AF 4   Cjun   34.94   0.165           C1   Blood 1   Cjun   35.39   0.218           C2   Blood 2   MN1TEL   34.37   0.281                                         C3   Blood 3   CRE   Undetermined                                             C4   Blood 5   Cjun   36.35   0.172           C5   Blood 6   MN1TEL   34.96   0.634           C6   Blood 7   CRE   37.76   0.556           C7   Blood 9   Cjun   33.61   0.069           C8   Blood 10   MN1TEL   34.3   0.734           C9   Blood 11   CRE   32.9   0.6                                         C10   AF 2   MHV   Undetermined               C11   AF 4   MHV   Undetermined                                         D1   Blood 1   Cjun   35.08   0.218           D2   Blood 2   MN1TEL   34.77   0.281           D3   Blood 3   CRE   39.09           D4   Blood 5   Cjun   36.6   0.172           D5   Blood 6   MN1TEL   34.06   0.634           D6   Blood 7   CRE   38.55   0.556           D7   Blood 9   Cjun   33.71   0.069           D8   Blood 10   MN1TEL   33.26   0.734           D9   Blood 11   CRE   33.74   0.6                                         D10   AF 2   MHV   Undetermined               D11   AF 4   MHV   Undetermined                                         E1   Blood 1   MN1TEL   33.7   0.131                                         E2   Blood 2   CRE   Undetermined                                             E3   Blood 4   Cjun   37.7   0.252           E4   Blood 5   MN1TEL   35.48   1.053           E5   Blood 6   CRE   31.84   0.03           E6   Blood 8   Cjun   34.57   0.13           E7   Blood 9   MN1TEL   32.45   0.111                                         E8   Blood 10   CRE   Undetermined                                             E9   AF 1   Cjun   39.35   0.278           E10   AF 3   Cjun   33.75   0.213           E11   BF 1   Cjun   28.14   0.048           F1   Blood 1   MN1TEL   33.52   0.131                                         F2   Blood 2   CRE   Undetermined                                             F3   Blood 4   Cjun   38.06   0.252           F4   Blood 5   MN1TEL   36.97   1.053           F5   Blood 6   CRE   31.88   0.03           F6   Blood 8   Cjun   34.75   0.13           F7   Blood 9   MN1TEL   32.29   0.111                                         F8   Blood 10   CRE   Undetermined                                             F9   AF 1   Cjun   38.96   0.278           F10   AF 3   Cjun   34.05   0.213           F11   BF 1   Cjun   28.21   0.048                                         G1   Blood 1   CRE   Undetermined                                             G2   Blood 3   Cjun   34.52   0.041           G3   Blood 4   MN1TEL   36.02   0.284           G4   Blood 5   CRE   38.12   0.071           G5   Blood 7   Cjun   34.69   0.387           G6   Blood 8   MN1TEL   33.29   0.302           G7   Blood 9   CRE   37.75           G8   Blood 11   Cjun   36.57   0.057                                         G9   AF 1   MHV   Undetermined               G10   AF 3   MHV   Undetermined           G11   BF 1   MHV   Undetermined           H1   Blood 1   CRE   Undetermined                                         H2   Blood 3   Cjun   34.46   0.041           H3   Blood 4   MN1TEL   35.62   0.284           H4   Blood 5   CRE   38.02   0.071           H5   Blood 7   Cjun   35.24   0.387           H6   Blood 8   MN1TEL   33.72   0.302                                         H7   Blood 9   CRE   Undetermined                                             H8   Blood 11   Cjun   36.65   0.057                                         H9   AF 1   MHV   Undetermined               H10   AF 3   MHV   Undetermined           H11   BF 1   MHV   Undetermined                      
 
      The screening results are transmitted to the remote user  1  within twenty-four hours of receiving the sample at the screening laboratory  20 .  
     Example 3  
     MHV (RNA Virus) Screening  
      Biomatter in the form of fecal swabs from mice is submitted via FedEx® (Memphis, Tenn.) overnight delivery. Each sample occupies one well of a 96 source well container  2 . The remote user  1  provides the genetic line identification  84 . The genetic line in this example has been previously associated by the remote user  1  with the designated genetic sequence for MHV (SEQ ID NO. 34). Samples are counted and 250 μl of SV Lysis reagent (Promega Corporation, Madison Wis., # Z305X) is added to each sample well of the source well container  2 . The source well container  2  is then vortexed to homogenize the samples. Next, the source well container  2  two is spun in a centrifuge for one minute.  
      The source well container  2  is then placed back on the deck of the Tecan Genesis Workstation® (Research Triangle Park, N.C.). Once all of the samples are transferred to the primary master well plate, the well plate is moved to the deck of the Isolation/Purification Station  94 .  
      One hundred and twelve microliters of lysis reagent (Promega Corporation #Z305X) are added to each sample. Thirty microliters of magnetic particles (Promega Corporation A220X) are added to the wells of a 384 destination well plate (Fisher Scientific #NC9134044). The well plate is moved into a magnetic field and the packing oil supernatant is aspirated off the particle bed. The liquid handler aspirates 100 μl of each sample liquid fecal biomatter sample and dispenses it into the 384 primary master well container, mixing the samples and particles. The particles are allowed to incubate at room temperature for three minutes with a sufficient amount of chaotropic salt to cover the particles. The primary master well container is then moved into a magnetic field where the magnetic particles are drawn to the bottom of each well. The supernatant are then aspirated and discarded. The primary master well container is then moved out of the magnetic field. Next, 150 μl of 95% ethanol is added. The primary master well container is moved into the magnetic field and the ethanol supernatant is aspirated off the bead bed. Then, the primary master well container is placed on a 384 tip dryer for one minute. Then the primary master well container is moved back to the deck of the Isolation/Purification Station  94  and 50 μl of DNase solution (Promega Corporation, Yellow Core Buffer #Z317D, MnCl 2  # Z318D and DNase # Z358A) is prepared according to Promega Technical Bulletin 328 and added to each sample and incubated at room temperature for 15 minutes. Next, 100 μl of stop buffer (Promega Corporation, DNase Stop #Z312D) is added and incubated for two minutes at room temperature. Two ethanol washes are done as described above. The primary master well container is then placed back on the dryer for two minutes. Finally, 60 μl Ambion&#39;s (Houston, Tex.) nuclease free water (catalog #B9934) is added to each well of the primary master well container. The elution solution is heated to 95° C. The primary master well container is then moved into the magnetic field and 50 μl of DNA was transferred to a 384 well optical storage plate (Fisher Scientific, #08-772136) for optical density analysis.  
      An A 260  reading of the storage plate read is performed with a Tecan Genios Spectrometer. This reading showed nucleic acid is present at the desired standard concentration of 0.2 O.D. units, but a range of 0.1 to 0.5 O.D. units is acceptable.  
      The plate with the isolated RNA is moved to the deck of a Tecan Freedom Workstation; reverse transcriptase-PCR mixture and Ambion water was placed on the deck as well as a 384 optical well plate (Applied Biosystems (Foster City, Calif.) catalog #4309849)). The reverse transcriptase-PCR mixture is made with TAQ-Man® EZ RT-PCR Kit (Applied Biosystems, catalog #N808-0236). The Tecan Genesis adds the reagents together in the ABI 7900 384 Well Optical Plate. The plate is then sealed with optical sealing tape (ABI, #4311971). The samples are incubated for two minutes at 50° C., thirty minutes at 60° C. and five minutes at 95° C. The plate is then thermocycled for twenty seconds at 94° C. and one minute at 62° C., for forty cycles. The results are shown in Table 6.  
                               TABLE 6                               Designated                   Sample   Genetic       Std. Dev.       Well   Name   Sequence   CT   CT                  A1   1 + Full   MHV   27.15   0.408       A2   1 + ¾   MHV   27.64   0.474       A3   1 + ½   MHV   28.41   0.226       A4   1 + ¼   MHV   32.5    1.917                                 A5   Water Full   MHV   Undetermined                                     B1   1 + Full   MHV   26.57   0.408       B2   1 + ¾   MHV   26.97   0.474       B3   1 + ½   MHV   28.09   0.226       B4   1 + ¼   MHV   29.79   1.917                                 B5   Water Full   MHV   Undetermined                                     C1   2 + Full   MHV   24.03   0.033       C2   2 + ¾   MHV   24.41   0.385       C3   2 + ½   MHV   24.86   0.252       C4   2 + ¼   MHV   26.21   0.273                                 C5   Water ¾   MHV   Undetermined                                     D1   2 + Full   MHV   23.98   0.033       D2   2 + ¾   MHV   23.87   0.385       D3   2 + ½   MHV   24.51   0.252       D4   2 + ¼   MHV   25.83   0.273                  
 
      The screening results are transmitted to the remote user  1  within twenty-four hours of receiving the sample at the screening laboratory  20 .  
     Example 4  
     Human Swab Screening  
      MasterAmp Nylon Buccal Swabs (MB030BR Epincentre, Madison, Wis.), are used to collect DNA samples from the oral cavities of human. The swabs tips were removed and placed in individual wells of a VWR-DYNBL deep 96 well plate. Four biological samples in the form of a frozen swabs are submitted via FedEx (Memphis, Tenn.) overnight delivery to the screening laboratory  20  from the remote user  1 . Each sample occupies one well of a 96-well source well container.  
      The bioinformatics for the human screening had previously been performed by Applied Biosystems. The AmpFLSTR® PCR Amplification Kit amplifies nine tetranucleotide STR loci and the Amelogenin locus in a single reaction tube. The microsatellites that are amplified include D3S1358, D5S818, D7S820, D8S1179, D13S317, D18S51, D21S11, FGA, and vWA. Additionally, the amelogenin locus is used for gender identification. The bioinformatics and primer sets for Applied Biosystem&#39;s AmpFLSTR® Profiler Plus® PCR Amplification Kit is proprietary, however, the kit performs to a standard based upon the TWGDAM recommended guidelines. (Technical Working Group on DNA Analysis Methods. 1995. Guidelines for a Quality Assurance Program for DNA Analysis. Crime Lab Digest 22:21-43).  
      A lysis reagent such Nuclei Lysing Solution (Promega Corporation, Madison, Wis. A7943) per sample) is gently poured into a 25 ml trough or reservoir and is placed on the deck of a Tecan Genesis Workstation (Research Triangle Park, N.C.). The liquid handler dispenses 150 μl of the lysis reagent in to each sample well of the source well container  2 . The well plate is resealed and placed on a vortex for 10 minutes. The well plate is then placed back on the deck of the Tecan Genesis Workstation (Research Triangle Park, N.C.). The liquid handler aspirates 50 μl of each sample and dispenses it in to a 384 well primary master well container (Fisher Scientific #NC9134044). Once all of the samples are transferred, the primary master well container is moved to the deck of the Isolation Station Purification Station  94 .  
      One-hundred and twelve microliters of SV Lysis reagent (Promega Corporation, Madison WI, # Z305X) a chaotropic salt are added to each sample. Next, 13 μl of magnetic particles (Promega Corporation, #A220X) are added and the well components are mixed. The well plate is then moved into the magnetic field of a magnet where the magnetic particles are drawn to the bottom of each well. The supernatant is then aspirated and discarded. The well plate is moved out of the magnetic field and 95 μl of SV Lysis reagent is added to each well and mixed. The well plate is then moved into the magnetic field and the supernatant is drawn off and discarded. This washing process is repeated two additional times. Next, the samples are washed four times in 130 μl of 95% ethanol as described above. After the fourth ethanol wash, the microwell container is placed on a 384 tip dryer for 11 minutes. Then the microwell container is moved back to the deck of the Isolation Station Purification Station  94  and 155 μl of Ambion&#39;s (Houston, Tex.) nuclease free water (catalog #B9934) is added to each well at room temperature. The plate is then moved into the magnetic field and 50 μl of DNA elution is transferred to a 384 well optical storage plate (Fisher Scientific, #08-772136) for optical density analysis. An A 260  reading of the storage plate read is performed with a Tecan Genios Spectrometer (Research Triangle Park, N.C.). This reading shows nucleic acid is present at the desired concentration of 0.2 O.D. units, but a range of 0.1 to 0.5 OD units is acceptable.  
      The primary master wellplate with the isolated DNA is moved to the deck of a Tecan Freedom Workstation. The AmpFLSTR® PCR Master Mix, AmpFLSTR® Profiler Plus® Primer Set and Taq DNA polymerase and Ambion water are placed on the deck as well. The final PCR mixture is made of 1× AmpFLSTR® PCR Master Mix, 1× AmpFLSTR® Profiler Plus® Primer Set (30 μl) and 40% isolated DNA (20 μl). The Tecan Genesis added the reagents together in the 384 Well PCR Plate. The plate is then sealed with optical sealing tape (ABI, #4311971).  
      The samples are then placed in an Applied Biosystems SDS 7000. A standard PCR protocol is followed by heating the samples to 95° C. for 11 minutes, followed by thermally cycling the samples 28 times between 94° C. for one minute, 59° C. for one minute and 72° C. for one minute. The thermal cycling is followed by a final extension step of 60° C. for 45 minutes. The final step is that 25° for an indefinite period of time.  
      The PCR wellplate with the isolated DNA is moved to the deck of a Tecan Freedom Workstation. The deionized formamide/GeneScan-500[ROX] internal Lane size standard (ABI, #401734) solution and the AmpFLSTR® Profiler Plus® allelic ladder are also loaded onto the deck of the Tecan Workstation. The Tecan Genesis added the 1.5 μl amplified PCR products to the 25 μl of AmpFLSTR® reagents in a 384 Well PCR Plate. Other well locations in the 384 Well PCR Plate were loaded with 1.5 μl AmpFLSTR® Profiler Plus® allelic ladder to and 25 μl of the AmpFLSTR® reagents.  
      The 384 plate is then placed into a sample tray and placed on the autosampler of the capillary electrophoresis machine. The ABI prism 3100 Genetic Analyzer performs the auto loading, capillary electrophoresis and data capture of the samples. On average, these results are transmitted to the remote user  1  within twenty-four hours of receiving the biological sample at the screening laboratory  20 . The screening results are shown in Table 7 and  FIGS. 22-25 .  
                               TABLE 7                           Human       Human           Locus (STR)   DNA 1   Human DNA 2   DNA 3   Human DNA 4                  D3S1358   14, 15   15, 18   14, 15   14, 17       vWA   17, 18   17   17, 18   18, 19       FGA   24, 26   22   21, 22   22, 23       D8S1179   13   14   9, 13   14       D21S11   30, 31.2   28, 32.2   29, 32.2   29.2, 30       D18S51   15, 19   13, 18   13   14, 15       D5S818   11, 13   9, 13   9.13   11       D13S317   8, 13   9, 12   12   8, 12       D7S820   11, 13   8, 11   9, 10    9       AMELOGENIN   X, X   X, Y   X, X   X, Y                  
 
      Although the present invention has been described and illustrated with respect to preferred embodiments and a preferred user thereof, it is not to be so limited since modifications and changes can be made therein which are within the full scope of the invention.