[Methods-and-reagents] 2nd strand with klenow

Tim HART Tim_HART at nea.gov.sg
Tue May 31 22:33:49 EST 2005


We have an RNA archive which I want to keep for as long as possible. I am 
trying to at least get a cDNA copy (with random primers) of the important 
RNA, so that we don't have to defrost and resuspend the RNA every time we 
want to use it. But I have been told that ssDNA isn't as stable as dsDNA. 
I am playing around with using Klenow to make the cDNA into dsDNA. Has 
anyone experience in this? Has anyone got a simple protocol for second 
strand synthesis? Is it possible to combine 1st and second strand 
synthesis into one reaction?

Thanks a lot
 
Tim Hart, PhD
Research Scientist
Environmental Health Institute
The National Environment Agency
DID          +65 67719113
Fax          +65 67778029
 
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