QC of qPCR replicates

Here we discuss use and processing of technical repeats

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QC of qPCR replicates

Postby greenwood » Mon Feb 13, 2017 11:22 am

We usually remove PCR replicates if they have a high standard deviation and repeat the measurements for the relevant samples and targets. I just realized that if I am analyzing my data with GenEx, it is not possible to merge the data I got from the repeat with the original data. We have a LightCycler480 and the plate setup of the original run and the repeat run are not the same.
I am not sure how we should proceed with qPCR replicates that have a high standard deviation instead of repeating the measurement. Just deleting the values doesn’t work as I am losing too much data then. Especially if it concerns reference gene assays, I could not normalize the samples.
Would you recommend to keep the Cq values and just use the mean of the qPCR replicates anyways? But what is the QC good for then? Or should we just repeat the whole run if a certain amount of replicates fails QC? Do you have any recommendations how to handle this issue?
Many thanks!
greenwood
 
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Re: QC of qPCR replicates

Postby MultiDAdmin » Wed Mar 22, 2017 2:18 pm

Hi

Please see our reply at the by the following link:

viewtopic.php?f=32&t=1009

BR
qpcrforum team
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Site Admin
 
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