Can the same convention pcr cyclic conditions be set for quantitative pcr?

23 July 2022 1 7K Report

I ran the following cyclic conditions in the conventional pcr and obtained good distinct bands (product size-114 bp). Please share your opinions whether I can go for the same set of conditions for qpcr as well. I am using takara TB Green Premix Ex Taq kit

Initial denaturation- 94° 2 min

35 cycles for the following set

94° 30s

50° 30s

72° 45s

Final extension- 72° 10 min

Pankaj Vinod Sharma

Dear Dikshita Saikia ,

DNA amplification in qPCR machine or normal machine is same. The only additional thing is detector dye in qPCR mix (i.e. TB GREEN in your case) that will not hinder your DNA amplification.

So, YES definitely you can go for same temperature conditions for qRT-PCR.

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