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NB transfer problem - (Jul/14/2001 )


I have a trivial problem, RNA is not easily transferred from the gel on the membrane during Northern blotting, by UV-illumination, I see that a majority remains in the gel, I use 1 % formaldehyde gels made with Sigamas general purpose agarose, transfer solution was 20 x SCC, transfer time was 1,5 days, and the membrane was Hypond N, anybody here who has an advice.


Try to use 0.5% formaldehyde gels instead 1%. It is strong enough to prevent RNA degradation and to denature RNA on the gel. If you can afford to buy vacume system from Pharmacia, it will increase the efficiency very much. You mentioned "RNA". Do you need to transfer rRNA completely also? If not, why bother? if only 28s and 18s are still on the gel?Julie