I previously ran three qPCR plates (20160121, 20160122, & 20160125) with 20 reps of each of the following copy numbers to determine the limit of detection (or, analytical sensitivity) of this qPCR assay: 30, 10, 3, 1
Additionally, I determined the average baseline threshold to set (580.5), based on eight qPCRs (see 20160128).
Performed calculations and determined limit of detection (defined as >95% of samples produced amplification) for this assay is three copies. The single copy sample amplified 88% of the time. This also means that our cycle threshold cut-off for this qPCR assay should be 37.9, as the mean Cq for three copies was 37.33, with a standard deviation of 0.53.
The data and calculations can be seen below (Google Sheet – scroll to the right to see calcs):
Google Sheet: RLOv_DNA_helicase_LoD_calcs