Efficiency differences between target and housekeeping gene - (May/10/2006 )
I was wondering what is an acceptable difference in the efficiency of amplification of a target and a housekeeping gene? Is there a method to calculate the results that can cope with large differences?
My target gene E is 81%, and my housekeeping gene is 95%. I have already optimised both for primers, MgCl, etc
If you use the formula developed by Pfaffl, Nucleic Acid Research, 29, 2001, you should be OK.