qPCR – RLOv DNA Helicase Standard Curve Check

Since the standard curve for this assay was a bit wonky at the low copy number end the last time I ran it, I made a fresh 1:10 dilution of the 3 copies component of the curve (100μL of 30 copy sample in 900μL TE).

Ran qPCR using the RLOv DNA helicase standard curve from 20151106.

All samples were run in duplicate.

Master mix calcs are here (Google Sheet): 20151223 – qPCR RLOv DNA Helicase Curve Check

Plate layout, cycling params, etc. can be found in the qPCR Report (see Results below).

Results:
qPCR Report (PDF): Sam_2015-12-23 13-57-01_CC009827.pdf
qPCR Data File (CFX96): Sam_2015-12-23 13-57-01_CC009827.pcrd

 

The efficiency & R^2 values look pretty solid, but the spacing between the 300 copy (Cq ~ 32 in the amplification plot) and the 30 copy (Cq ~ 34 in the amplification plot) samples is a bit too tight for my liking. Additionally, the reps for the 3 copy sample are very poor.

Will repeat to see if I can tighten things up…

One thought on “qPCR – RLOv DNA Helicase Standard Curve Check

  1. Pingback: qPCR – RLOv DNA Helicase Standard Curve Check Repeat | Sam's Notebook

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