10-18-11 In today's lab we analyzed our PCR products from last Thursday. We ran all 31 samples through gel electrophoresis and had some positive results. We found from running our B primers at the different annealing temperatures that the highest temp. that they can successfully work at is 58 degrees celcius. Our gel also showed banding around 1100 bp for three of our regular flanking primers and three of our biobrick primers. Banding at that size indicates that we may have successfully joined our two exon fragments. There was quite a bit of non-specific primer binding at most of the temperatures as well, so the gel was allowed to run an additional 30 minutes (had previously ran 30min before analysis) to get better resolution of the successful bands. We also found that 3 annealing temperatures worked very well for each of the primer sets (regular and biobrick), and these six lanes will be used to cut DNA from. (lanes 4,5,6,12,13,14). Temps ranging from 44-52 degrees celcius.
Axel is going to remove our gel once the additional 30 minutes is up and will also cut those bands at 1100 BP out of the gel, which will be used for DNA extraction during our next lab.
Written by Kim Lovik