What is the amount of RNA (adipose tissue origin) used for 1-step RT-qPCR?

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What is the best concentration of cDNA to be used for qPCR?

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Han Zhang

Hi Mikey, I once used 1-step RT-PCR kit and 50 ng of RNA works well, however it is not from adipose tissue. You need to optimize the amount of RNA.

Alex Young

The 25-40ng you suggested should be sufficient, I have done many qPCR assays with just 20 and had success. Good luck.

Sven Reister

Hi,

this is pretty much also dependent on your gene expression. For strong expressed gene also lower inputs like 1 or 5 ng total RNA are sufficient. I would not exceed the 20ng Alex has recommend except your gene of interest is very low expressed.

Sven