PCR Lab1 - Lionel Sims III Lab T.A Dr Darlene...

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Lionel Sims III 02/06/08 Lab T.A.: Dr. Darlene Campbell (Parts A and B) Determining Allelic Frequencies Using PCR Technology and Gel Electrophoresis Abstract This experiment was designed to provide understanding of and practice with PCR (polymerase chain reaction) technology. It was also a chance for each participating member to explore their own genotype and compare it to those of others in a sample population. I took a small amount of my own DNA, extrapolated from buccal cells with a chemical process involving saline, and proceeded to run them through a thermal cycler, Using PCR technology to exponentially increase the number of DNA molecules, the thermal cycler ran for hours. The sample was then dyed and put into an agarose gel for gel electrophoresis. The electrical charge of the nodes at opposite ends of the gel and the charge on the DNA molecules caused them to move through the gel and separate based on size and weight. Afterwards the results were obtained and more results were calculated from those. I expected the results to show a genotype similar to the majority of African Americans, but that wasn’t the case. Ultimately the data showed that I have a genotype of 27/34. It is not a very common one and especially so among African- Americans. The data also showed possible contamination in the gel in lane 1 of the wells which was designed to operate as an Identifier of contamination, however it seems to be miniscule enough to be negligible. Results I performed Gel Electrophoresis at 190 volts for 25 minutes with an allelic ladder for the D1S80 allele and measured the distance traveled by each band in millimeters on a printed ultraviolet image representation of the agarose gel after Gel Electrophoresis was complete. Each band on the allelic ladder was intended to represent a specific number of Variable number tandem repeats (VNTR’s). The number of VNTR’s allowed the calculations of the number of base pairs that each allele represented in the sample and
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This lab report was uploaded on 04/18/2008 for the course BIO G 106 taught by Professor Campbell,d. during the Spring '07 term at Cornell.

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PCR Lab1 - Lionel Sims III Lab T.A Dr Darlene...

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