When converting human brain total RNA to cDNA my results keep coming back incorrect. Is there something wrong with my standards?

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Zandisiwe Magwebu

Here is the standard curve

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.

i'm currently using cDNA that i converted from the total RNA.