Earlier today I made some cDNA from geoduck gonad RNA for use in this qPCR to test out the vitellogenin primers I designed on 20181129
I also used geoduck gDNA (162ng/uL; from 20170105) as a potential positive control, or as confirmation that these primers will not amplify gDNA.
Primers:
- 1712
- 1711
All qPCR reactions were run in duplicate. See qPCR Report (Results section below) for plate layout, cycling params, etc.
qPCR Master Mix calcs (Google Sheet):
Results
qPCR Report (PDF):
CFX Data File (PCRD):
Primers worked perfectly (although, expression comes up a bit later than I’d like)! Good amplification, single peak in melt curve, and no amplification in gDNA.
This also means I can use this cDNA as a positive control for future geoduck VTG qPCRs.
Key for plots below:
- Green = cDNA
- Blue = gDNA
- Red = No Template Controls (NTC)
AMPLIFCATION PLOT
MELT CURVE
