In today's lab, we took the PCR products from Tuesday's lab and conducted an agarose gel electrophoresis to analyze it. We did a gel electrophoresis in order to determine which annealing temperatures would be optimal for another, larger PCR. We prepared the gel and placed the different samples in the lanes. After running the machine, we took a picture of the results. We found that the highest temperature (59.9 degrees) did not show any bands so was not suitable for either of our primer pairs, the middle temperature (47.8 degrees) showed the brightest bands for the "Exon A" primers, and the lowest temperature (44.0 degrees) showed the brightest bands for the "Exon B" primers. We will use the middle temperature for the A primers and the lowest temperature for the B primers in the PCR next lab period. We plan on using a larger mix, equaling 50 microliters instead of the 20 microliters used in the first PCR, in order to use more of our template DNA (10 microliters instead of 4 microliters).
Written by: Megan Hughes