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
>MY E-MAIL ADDRESS IS NGUYEND at ALLEG.EDU. THIS IS VERY URGENT, BECAUSE I
>CANNOT CONTINUE MY RESEARCH UNTIL I SOLVE THIS PROBLEM.
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.
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