Help: Blotting DNA from acrylamide gel

Daniel Kim dkim at nmsu.edu
Wed Jul 19 10:01:18 EST 1995


In article <173DF8842S86.SBROWN at GAES.GRIFFIN.PEACHNET.EDU> SBROWN at GAES.GRIFFIN.PEACHNET.EDU (Stuart Brown) writes:
>This is a real stumper for me.  I need to make a Southern blot of some
>PCR products after separating them by size in a polyacrylamide gel.  I

(stuff omitted)

>because nothing that they have tried worked very well.  Come on, this has
>got to be possible!
> 
>Does anyone out there have any experience blotting DNA out of acrylamide?
>No reasonable suggestions sneered at.

Hello:

I have no experience with this thing, but I was wondering what membrane 
you were using?  I know that you need a high salt buffer for nucleic 
acids to adhere to some membranes (e.g. nitrocellulose), unlike proteins, 
so that's why Southerns are not done in TBE, but in 10 - 20 x SSC.

You might want to consider your membrane choice, as well as transfer 
conditions.

Daniel Kim




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