Hi,

I have to do qPCR experiments right now and keep on running into the same problem again and again with different samples.

The image shows the amplification curves of 3 technical replicates of the the same sample. Mastermix is the same (30uL) to which 20 uL of the same DNA from the same tube was added. Samples are mixed by pipetting up and down a few times.

I assume that the DNA quantity was very similar because the curves pass the threshold almost simultaneously and the linear phase is similar too.

What I don't understand is why they reach so different plateaus and how to interpret it. Therefore I am also not sure how to troubleshoot it.

Any help would me much appreciated.

Thanks a lot.