very low expression of a sample vs standard curve efficiency - (Jul/16/2010 )
I am currently working with a gene expression pattern under two different conditions.
I do get a Ct for my gene around 24, but I can not get a good efficiency on my stantard curve because I only get reading till a 1:50 dilution.
I use cDNA for my standard curve. Can I use genomic DNA for the standard curve? or there is something else I should do?
I have tried very low dilutions (1:5 1:10 1:15 1:20) but as expected the efficiency is not even close to 80%.
Thank you so much!
if your primers are detecting gDNA (no intron spanning primers) than you can use gDNA for the STD curve, but I would consider to design new primers to improve the efficiency.