This is a repeat of an earlier run from today, but with a different qPCR plate. Here’re master mix calcs (bottom half of page). Cycling params were as follows:

• 95C – 10min

40 Cycles of:

• 95C – 15s
• 55C – 30s
• 72C – 1m

Melt curve.

Results:

All samples amplified and showed a proper dissociation curve. However, it does look like there’s a spread of ~3 Cts across the plate. This is not good, as this is the equivalent of ~10-fold difference in gene expression. Will repeat again and see if specific wells show the same relative differences in Cts.