In today's lab, we ran a gel electrophoresis on our PCR products from last lab. The PCR attempted to connect the two exons of our DNA sample into one complete strand at around 1100 bp. The results of the gel showed that banding was present at around 300 bp and 800 bp at all of the annealing temperatures used in PCR. The PCR was not successful we suspect because the template DNA sample was not clean enough, and still contained the primers used to cut the DNA into 2 pieces.
We then cleaned our PCR product using the GeneJet Purification kit protocol, and set up a new PCR reaction using the purified PCR in 8 different annealing temperatures. We mixed 8 tubes with our flanking primers, the biobrick primers, and the second set of primers (the "B" exon primers). Another tube was made with our first set of primers (the "A" exon primers), and created controls for all of the above. We are expecting the purified PCR will come out with better results, with one DNA strand of approximately 1100 bp. Next lab will will run a gel electrophoresis to view and analyze our results of the PCR. We hope to see which temperature(s) is/are optimal, and at what temperature(s) will no longer work with our primers.
Written By: Megan Hughes