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RNA concentration to start RT reaction


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#1 virusman2

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Posted 12 June 2013 - 08:38 AM

I am working with a variety of tissue samples to detect some viruses.
measuring RNA concentration in the extracts reveals a wide variation, with some samples have as low RNA as 50ng/ul.
i am wondering if there is a minimal limit of the starting RNA in the RT reaction, so that one can avoid testing useless samples.

Edited by virusman2, 12 June 2013 - 08:39 AM.


#2 doxorubicin

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Posted 12 June 2013 - 11:32 AM

I'm sure your RT enzyme has some recommendations in the protocol. For example, SuperScript II RT suggests "A 20-μL reaction volume can be used for 1 ng–5 μg of total RNA or 1–500 ng of mRNA." For this protocol you would be able to add 500ng per reaction (i.e. 10ul* 50ng/ul)....nicely within the suggested range.




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