35S in RNase Protection?

Eric Hanson hansone at bcc.orst.edu
Thu Jun 22 04:32:26 EST 1995


Has anyone had the experience of using 35S labeled RNA for RNase protection 
experiments? I was thinking of labeling RNA off an Sp6 or T7 promoter with 
35SUTP and using this as a probe for transcripts made from  in vitro 
transcription reaction using nuclear extracts and my promoter of 
interest.Will this be sensitive enough in this type of RNase protection 
experiment (of couse it depends on the amount of in vitro transcripts)? Any 
other drawbacks I should be aware of? I am thinking of this as an 
alternative to a 32P-labeled RNA probe since the 35S-labeled probe will 
last much longer and be more stable. I was thinking of using DIG but have 
convinced this is not the way to go.

Thanks for your help.

Eric 
hansone at bcc.orst.edu




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