How to check if DNA is transfered

Charles Rivers RIVERS at IPRSH.DOM.UAB.EDU
Tue Mar 23 19:56:20 EST 1993


Does anyone know of a way to check and see if any DNA has transfered to a
nylon membrane after a neutral, capillary southern blot?  I know that you
can check the collapsed gel, but what about the membrane?  Here's the
problem:  I tried to transfer 5 micrograms of digested DNA in a 0.75% agarose
gel.  I noticed that my gel had collapsed VERY early--like within an hour;
this made me very nervous, but I let the transfer continue.  When the transfer
was complete (from time), I restained the gel to see if the transfer was
indeed complete--it wasn't.  As a matter of fact, it looked like nothing
transfered at all.  Being spring and all, I've gotten lazy and don't want
to repeat an entire digest series.  I just want to be able to check the 
membrane and see if any of the DNA transfered.  I seem to recall that there
is a way of staining the nylon membranes with methylene blue to see if any
DNA is bound, rather than wasting time with a legnthy (sp?) hybridization
and detection.  Any information would be very appreciated.

Thanks,
Charles Rivers
Rivers at iprsh.dom.uab.edu



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