Selective removal of standards post qPCR Run - (Jun/05/2013 )
I was curious as to the rules for removing values from a standard reaction in a qPCR run. In qPCR runs where the effciency is just outside the 90-110% range and removing one standard dilution was that non linear relative to the other standards or the removal of one replicate within standard brings the run back within the 90-100% range, what is the rule for doing so? In this case the test being done is absolute quantification. Obiviously the data output will be altered when standards are adjusted, but unless subtle changes are being examined does this really matter?
Thank you very much for any information that can be provided. I want to make sure that any steps taken are within the norms of qPCR experimentation, and I feel that this is a good place to get that feedback.
I do this. I always change the threshold manually, kicking out any outliers. Just make sure you adjust your control thresholds to match your experimental samples.