qPCR assay Triplicate Ct SD
Entering edit mode
7 months ago
Maria17 ▴ 20

Hi All,

Currently, I'm working on a qPCR assay. Since all targets are done in triplicate, I don't know the acceptable range of Ct Value Standard Deviation between replicates. Most of my targets have a CT SD less than 1, but some have a CT SD between 1-2. What should I do with those targets? Should I discard them or should I leave them as they are?

Any recommendations or solutions would be appreciated.



miRNa qPCR RT-qPCR • 364 views
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There's no strict or generally accepted cutoff, but I probably wouldn't use this data for final reporting in a publication. I'd look into why this one target has an unusually high CT SD and work to alleviate that issue first. I suggest freezing the cDNA/RNA you're analyzing, as its probably some problem related to primer efficiency, reaction conditions, low starting copy number, etc.. which you could optimize and then re-run your initial targets with hopefully lower CT SD

Entering edit mode

Thank you for your response. How about the CT values for the two plasma samples (tubes) from the same patient?

In some cases, I have to measure the same miRNA target in several tubes from the same patient, but some of the target values are different. According to what I understand, the target CT values in both tubes from the same patient should be identical or similar.

It would be great to have your response to this issue.

Regards Maria


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