Ran PCR using primers Cg_COX1/2_qPCR_F, Cg_COX1_qPCR_R, Cg_COX2_454align1_R (SR IDs: 1192, 1191, 1190; respectively). Template was pooled cDNA from 20110311 of various C.gigas tissues. These reactions will verify (sort of) if we have both isoforms present in the PCR performed earlier today, prior to cloning. Master mix calcs and cycling params are here.
Results:
(http://eagle.fish.washington.edu/Arabidopsis/20111007-01.JPG)
Lane 1: Hyperladder I (Bioline)
Lane 2: COX1/PGS1 primer set
Lane 3: COX1/PGS1 primer set NTC
Lane 4: COX2/PGS2 primer set
Lane 5: COX2/PGS2 primer set NTC
NTCs are clean for both primer sets. We see bands of the expected size for both primer sets. Additionally, we see lower expression in COX2/PGS2, as we observed in our previous qPCR reactions with these primer sets. Will clone the large fragment that was PCR’ed/purified from earlier today.