What should be the optimum product size for getting precise amplification for expression of a particular gene?

01 January 1970 1 1K Report

In my qPCR, I have designed primers for 38 genes of Glucosinolate biosynthesis in cabbage. Where, my primers product sizes of those from 110 bp to 350 bp. However, in case of two genes, I have to design 500 bp for getting good amplification curve. Am I in right tract or not?

Diego Batista

Dear Ujjal Kumar Nath ,

The problem with longer amplicons is that will increase the chances of detection something not desired, which may entirely lie outside to your target. So even with a good amplification curve I suggest avoiding to use > 200bp amplicons in plant essays.

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