Bio 13 Lab Report

In this three-week long experiment conducted in the Bio 13 Lab, we were able to analyze a single nucleotide polymorphism (SNP) in our own genomic DNA and then determine our genotype at this specific SNP. In week one, we extracted genomic DNA from our cheek cells with swabs and prepared our DNA for PCR (Polymerase Chain Reaction) that would amplify the region with the intended SNP of interest. After one week and after the PCR was run outside of the lab section, the resulting PCR product was purified and treated with restriction enzyme Ahdl in order to prepare for the final analysis of our genotypes. In the third and final week of the project, we analyzed our PCR products by means of agarose gel electrophoresis. By the conclusion of the experiment, we had completed the analysis at the SNP of interest and determined our genotypes for this SNP. The first experiment involved in this four-week project was the extraction and

Once the program was run, the machine began its thirty-five PCR cycles. Each step in these three step cycles is defined by a specific time span and temperature. However, before the first cycle begins, initial DNA denaturation occurred for five minutes at a temperature of 95 oC in order to ensure that all the DNA had become single-stranded. After this initial denaturation, the thirty-five PCR cycles could begin, first with thirty seconds of DNA denaturation at 95 oC, then followed by a drop of temperature to 58 oC for thirty seconds for the primers to anneal to its target sequence on the template DNA, and finally thirty seconds at 72 oC for primer extension in which DNA synthesis is carried out by Taq Polymerase. After the thirty-five cycles were complete, there was an additional five minutes at 72 oC for the final extension of the DNA to ensure that all synthesis had been completed. The completed PCR product is then held at 12 oC to preserve it until removed from the