Hello,
I would like to ask a question. I want to do PCR from a large amount of sample, but it does not fit on my plate. This means that in order to test them all, I have to put them on more plates, which means doing at least two PCR runs. Then I would like to compare each sample with each other. My question is. Is it necessary to have a housekeeping gene in every PCR run? Or it is enough to have it only in the first run and relate the samples from other PCR runs to this housekeeping gene.
Thank you for answer.
Yes it is necessary to include a housekeeping gene for each plate, and calculate expression values based on that. Cannot extrapolate values for housekeeping genes as each PCR run is different due to small pipetting errors, machine readings, reagent thaw/freeze cycles etc
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