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very low expression of a sample vs standard curve efficiency

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#1 boristhecat



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Posted 16 July 2010 - 10:18 AM


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!


#2 tea-test



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Posted 18 July 2010 - 11:11 PM

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.
tea-test: The artist formerly known as Ned Land

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