I was wondering what is a good amount of RNA to use to get good ct values? I have been using 50ng/ul (total 250ng) per reaction and I am getting slightly high ct values. Any advice would be appreciated.
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jerryshelly1
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Posted 06 February 2013 - 11:57 AM
What cDNA kit are you using?
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Wek
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Posted 17 February 2013 - 11:23 PM
jerryshelly1, on 06 February 2013 - 11:57 AM, said:
What cDNA kit are you using?
Sorry for the late response.
Hmmm.. I don't really know the kit I am using to make cDNA. All the reagents I use were taken out of the package. I remember I use random hexamers and denature the RNA then add rnase, dntps, a buffer, and ssi (i think that's the name).
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jerryshelly1
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Posted 18 February 2013 - 08:38 AM
Just double check the recommended amount of RNA (ug) that needs to be added with each reaction. If available, use two or three different specs from a couple different labs to confirm your RNA concentration. That can be the biggest contributing factor.
