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Amount of RNA (ng) for good ct values.


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3 replies to this topic

#1 Wek

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Posted 06 February 2013 - 10:34 AM

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.

#2 jerryshelly1

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Posted 06 February 2013 - 11:57 AM

What cDNA kit are you using?

#3 Wek

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Posted 17 February 2013 - 11:23 PM

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).

#4 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.




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