DNA Sequencing

    Although DNA sequencing is a great way to extract information based on genotype of a specific organism, it is not the only way.  In this experiment, genotypic information about the dogs was gathered using a combination of polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and derived cleaved amplified polymorphic sequences (dCAPs). PCR is a method of DNA amplification that can produce an exponential number of copies of a specific sequence. PCR is run in a thermocycler in a tube containing taq polymerase, dNTPs and a primer specific to the section of DNA to be amplified. The end product of PCR is millions of copies of the desired DNA sequence. The amplified signal is useful for reliable repeated assays of that target sequence. For this lab, the product of PCR was used in dCAPs to figure out the genotype of the dogs at a gene. dCAPs is a derived version of RFLP that can be used for SNPs without a natural restriction site. Both RFLP and dCAPs use a restriction site containing a desired SNP to determine the genotype at a gene, by cutting the sequence with a restriction enzyme and running the product(s) of the digestion on a gel