Northern Blot problem

Kenneth Y. Ilio you at somehost.somedomain
Thu Aug 24 17:20:32 EST 1995


In article <01HUC4ZQAK8I8Y629G at VAX.CS.HSCSYR.EDU>, Karen.Straube-West 
says:
>
>Hi Bionetters,
>
>We are suddenly having difficulty with our Northern blots using total 
RNA.
>The appropriate bands are present, but they are smeary. Even the rRNA on
>the gel looks smeary. It isn't bad RNA because it looks fine on a
>non-denaturing gel.
>
>We've checked, and the pH of our formaldehyde is about 3.2. Our protocol
>says it should be >4.0. Question: Could this be the problem? If so, can 
we
>raise the pH of our formaldehyde or should we buy new stuff? I'd 
appreciate
>any advice.
>
>Thanks,
>
>Dr. Karen Straube-West
>SUNY Health Science Center
>750 East Adams Street
>Syracuse, NY 13210
>
>
>
I have done Northern blots extensively in the past. Also experienced some 
problems with the blot. However, formaldehyde does not appear to be a 
problem as all formaldehyde used in our laboratory have pH consistently 
less than 4.0. My problems went away when I began running the gel for 
longer time. I hope this will help you.



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