Quant of InVitro synthesized RNA

Zeiler, Brian M'bio BrianZ at MICROBIO.LIFESCI.UCLA.EDU
Fri Aug 25 17:12:26 EST 1995


This was posted today:

>I need to quantitate pmoles of T7 synthesized 32P-labelled RNA, as well as
>unlabelled invitro synthesized RNA. What are the preferred methods, do you
>gel purify the RNA, or NH4Ac ppt from the synthesis rxn? thanks

You need to get rid of unincorporated counts in order to accurately 
quantitate your RNAs.  In my hands, gel purification is the best method and 
the only one I trust.  I use tritium labelled RNA as "unlabelled" and run it 
next hot stuff so that I can cut it out of a gel.

I will be happy to share with you my entire protocol if you will send me 
your e-mail address.

Brian Zeiler
brianz at microbio.lifesci.ucla.edu



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