alkali vs non-alkali transfers

Ross S Feldberg rfeldber at emerald.tufts.edu
Fri Jul 18 08:10:20 EST 1997


Hi;

 I am setting up Northern blots and was wondering about optimal transfer
conditions. We are using downward blotting onto charged nylon filters with
an alkaline transfer solution. The protocol we are following calls for
only a 60 min transfer in order to minimize hydrolysis of the RNA.
However, I am not sure if we are getting good transfer in this time frame. 

I was wondering if anyone uses non-alkaline transfer medium or whether
hydrolyis is enough of a problem to worry about. How long do folks
generally transfer for?  (I ask because our initial efforts with an actin
probe gave a pretty weak signal).


Thanks

Ross Feldberg
Biology
Tufts Univ.
rfeldber at Tufts.edu




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