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This is my answer, but wait for other people on this forum to answer too:
if this is only for relative comparision then just calculate the delta Ct as always. unless you want to do quantification and draw standard curve? then to me it is risky to calculate, because there is a high chance of error. However, you can try reverse ratio. I mean for example if your Ct is 15 for a 1/8 sample, and you want to know the possible Ct for 1/6 you can try x=15*0.125/0.16=11.71. do this for any GAPDH sample.
I would recommend having same ratio cDNA for next experiment.
I used RotoGene software. However I don't remember if the software allowed adding the standard curve manually?!
see this handbook too:
www.qiagen.com/literature/render.aspx?id=23490
#131245 qPCR normalization of reference gene
Posted
on 18 March 2012 - 09:20 PM
