Southern blotting from acrylamide gels?

S.K.Green S.K.Green at shef.ac.uk
Mon Oct 16 10:42:14 EST 1995


JJAM <U55355 at uicvm.uic.edu> wrote:
>Does anyone out there do Southerns from PAGE gels?  No one I know has ever done
>it or heard of it.  Can it be accomplished using capillary transfer or is
>electroelution necessary?  Any advice would be most appreciated.


The answer is YES.  I used to do this by electroblotting, but you could 
probably use a vacuum blotter as well.  This is my method.

1.  Run gel
2.  Wash in 1xTBE
3.  Set up a blot sandwich and blot in 0.5xTBE, at 0.5A, with cooling, 
for 3-12 hours.
4.  wash membrane in 1xTBE for 10min
5.  denature DNA by resting membrane on top of 0.4M NaOH saturated 
whatman 3MM for 10 min.
6.  Prehybridise and Hybridise your membrane as normal

Good luck

Sue Green.




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