Help needed on blotting of DNA from native polyacrylamide gels

Dennis J. Templeton djt2 at po.CWRU.Edu
Tue Feb 16 14:38:55 EST 1993


In a previous article, nguyend at carr106.alleg.edu (Doan Nguyen) says:

>
>I NEED TO KNOW IF ANYONE HAS ATTEMPTED TO BLOT DNA FROM NATIVE  
>POLYACRYLAMIDE GELS.  CAN IT BE DONE BY CAPILLARY ACTION OR SHOULD IT BE  
>DONE BY ELECTROBLOTTING?  
>
>ANY INFORMATION WOULD BE GREATLY APPRECIATED.  PLEASE GIVE PROTOCOLS IF  
>POSSIBLE.
>
>MY E-MAIL ADDRESS IS NGUYEND at ALLEG.EDU.  THIS IS VERY URGENT, BECAUSE I  
>CANNOT CONTINUE MY RESEARCH UNTIL I SOLVE THIS PROBLEM.
>
>THANK YOU.
>
>DOAN NGUYEN
>

I had very good luck some years ago electroblotting onto nylon membranes in
a tank style electroblotter like one would for a western.

After blotting, I laid the membranes on a sponge full of 0.4M NaOH to fix
and denature the DNA.

As I recall, I used TBE running buffer at 0.05x (about 25 mM?) but I would
check your amperage first.

I don't think capillary blotting would work well.

good luck

dennis



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