How to standardize RT-PCR standard curve?

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Nathaniel Wade Mcgregor

Dear Aruna

Which methodology are you using? The 2002 method by Cawthon relies on relative quantification independent of an absolute standard. I would suggest reviewing the modification of this method by O'Callaghan and Fenech, 2011. They incorporate the use of a quantitative standard for absolute and relative quantification of telomere length.

Good luck!

Aruna Barade

Right now, I am trying to obtain standard curve using TEL primers. The peaks are stacked in one region and the points of different standard concentration are all aligned towards a particular region on the standard graph.