Hi people,
I'm struggling with a problem in my qPCRs. In every plate I have some wells (and each time different ones) that result in Ct values that do not make sense (e.g. Ct 4 when it should be around 22). The other wells are fine. The curious thing is that I have everything in triplicates and of the three wells it is usually just one that fails ! I have had this problem for a while now and obviously tried to find out whether it is some contaminated stock, but replacing everything did not help. Also, since I do not see a pattern, I cannot pinpoint it to a certain material or procedure step - I have had this phenomenon with different primers, different targets...
But it's the fact that the corresponding technical triplicates are fine which strikes me most, because this means the problem is not explainable by contamination of a certain reagent.
Any ideas ? I simply can't find an explanation for this phenomenon. Thanks...