This is an exact repeat of the PCR from yesterday, due to inconsistencies between repeated PCRs from yesterday and earlier today. Here’s yesterday’s workup. Samples were stored @ 4C O/N after completion of PCR. Gell was run on 20091217.
Results: This is getting embarrassing. Opsin results are same as yesterday (good!), but Rhodopsin results are slightly different than yesterday’s AND the day before yesterday’s results!
Gel Loading (from left to right):
Opsin Primers (lanes 2-10), Rhodopsin Primers (same loading order, lanes 12-20)
1 - 100bp ladder
2 - retina
3 - fin
4 - 4th arm
5 - dorsal mantle center
6 - dorsal mantle side
7 - ventral mantle center
8 - ventral mantle side
9 - H2O
10 - H2O
Opsin Primers
We see a band in the retina, fin, 4th arm, & ventral mantle center samples as we did yesterday. This replicates yesterday’s PCR results, which is good. Negative controls are clean. Bands will be excised and stored @-20C in Sam’s “Purified Inserts” box for eventual sequencing. NOTE: This gel was run on 20091217 and the tubes are dated as such!
Rhodopsin Primers
A single band is present in the retina, dorsal mantle center and ventral mantle center samples. We see TWO bands in the fin sample. These results differ from yesterday’s in that yesterday, a single band was present ONLY in the retina and fin samples. Two days ago there was a single band in each of the retina and fin samples and two bands in the ventral mantle center sample. Bands will be excised and stored @-20C in Sam’s “Purified Inserts” box for eventual sequencing. NOTE: This gel was run on 20091217 and the tubes are dated as such!
I am going to repeat these PCRs again. I can’t stand the fact that I am getting such freaking inconsistent results in an extremely simple PCR. Aaargh!