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.
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):
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)