Sam’s Notebook: qPCR – C.gigas primer and gDNA tests with 18s and EF1 primers


The [qPCR I ran earlier today to check for residual gDNA in Ronit’s DNased RNA] turned out terribly, due to a combination of bad primers and, possibly, bad gDNA.

I tracked down some different primers for testing:

  • Cg_18s_1644_F (SRID 1168)
  • Cg_18s_1750_R (SRID 1169)
  • EF1_qPCR_5′ (SRID 309)
  • EF1_qPCR_3′ (SRID 310)

In addition to BB15 from 20090519, I decided to test out BB16 from 20090519 as a positive control.

Samples were run on Roberts Lab CFX Connect (BioRad). All samples were run in duplicate. See qPCR Report (Results section) for plate layout, cycling params, etc.

qPCR master mix calcs (Google Sheet):