I keep getting incorrect results, please any suggestions? I prepared 1:10 dilution using human brain total RNA. I've converted to cDNA and my results keep coming back incorrect and the efficiency is ridiculously high (12, 734.242), slope = -0.474 and correlation coefficient = 0.212.
Jo Vandesompele
I recommend making a cDNA dilution series (instead of RNA). Further, either your data-analysis settings are not correct and/or the input is too high, as the first curves (lowest Cq) are not sigmoidal. All qPCR amplification curves should be parallel.
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