840:153g:Projects/project14/2011/09/27

9/27/11 In today's lab we assembled a PCR reaction to be run overnight. We used the leaf DNA sample based on the results from the gel ran on 9/22/11. That gel indicated a higher concentration of DNA in the leaf samples based on brightness and thickness of the DNA bands when compared to the rose petal DNA samples. A master PCR mix was prepared for the "A" primers, and a separate masteer mix was prepared for the "B" primers. The A primers will amplify our first exon, and the B primers will amplify our second exon. Three annealing temperatures were used for each set of primers (44.0, 47.8, 59.9 degrees celcius). A negative control was also prepared for each set of primers, each contained all essential PCR components, except for DNA. One positive control sample was prepared using DNA sample 1A7, and two primers (VF2,VR). A negative control was also prepared for the positive control sample, which contained all essential PCR components, except DNA. The PCR reaction will be run overnight, placed in the freezer, and will be tested using gel electrophoresis during the next lab period on 9/29/11.

The gel will be run to determine if the PCR reaction worked, and if so, which annealing temperature works best to amplify our DNA. If there is time, we will set up the second PCR reaction based on gel results.

Written by Kim Lovik.