How to remove RNase from protein extract?

brett brett at BORCIM.WUSTL.EDU
Wed Dec 11 09:18:30 EST 1996


>Hello there:
>
>I am doing experiments on a kind of RNA binding protein. Unfortunately, I 
>found the RNAs I had synthesized were all degraded by the RNase exsits in 
>the Protein extract. I used  to add RNasin in the reaction chamber,but it 
>did not work. Can you tell me how to discard the RNase without interfering 
>with the acitvities of other proteins? 
>
>Any suggestion will be greatly appreciated!
>
>Toy

I hope you reduced your samples before adding the RNasin. It needs 5mM DTT for
it to work, otherwise you may as well not add it at all. You could also try
inhibiting RNases with vanadyl ribonucleoside complexes. Or, if your protein
exhibits specificity for a particular RNA, why not dump in something like
tRNA?


Brett Lindenbach
    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu                             




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