We have been getting Ct values almost similar to a test sample in NTC wells. The melt curve also overlaps with the test samples. While running a gel, a band with a product size similar to the target gene is observed. We tried using new set of RNAse free water, cleaned the area, used filter guarded tips and also freshly diluted primers from the stock.
Yet this is happening for most of the genes.
Péter Gyarmati
Do you get the expected amplicon size? If no, there may be primer dimers causing your signals
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Musheer Aalam
Mostly your stock might be contaminated. Try your RTPCR with new set of stock primers.
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