Dear colleagues
I am about to choose reference gene for my qPCR experiments. I will do my measurements based on standard curve method. I have difference 2% (0.95-0.93) and want to know is it a good result or no. Could you please suggest what is the acceptable range of differences of standard curve efficiency between reference and target gene? Please suggest some papers.
Thank you in advance
Hi Hovsep,
The first question that I have for you is if you have duplicates or triplicates for each point on you standard curve? If you don't you should have. Do you have a an image from your standard curve that you can show?
Dear Erika, I have triplicates, but cannot show you any picture right now. Efficiency for reference gene curve is 0.95, is 0.93 ok for target gene?
You can use the Pfaffl correction to account for differences in efficiency between different primers.
http://www.gene-quantification.org/qpcr2004-pfaffl.pdf
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