I was trying different housekeeping genes in qPCR analysis and every combination of them gave me another result. When I choose one HK, two, or three, the results were completely different. I am confused because all of them are stable in my samples. (b2m, tbp, eif4a2; placenta tissues)
What is the best amount of HK genes? How to evaluate this data correctly?
I am a qPCR beginner :)
What did you mean by "all of them are stable in my samples"? Which house keeping gene to use may depend on the condition, tissue etc. Do you see stable high expression of your HK gene in your samples? What are the Ct values for your HK genes? Please provide more info.
3 HKGs would suffice. However, only after efficiency analysis should you choose your HKGs given that they all are behaving.
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