Blotting BIG gels (how??)
nigel at notmendel.Berkeley.EDU
Sat May 30 13:51:46 EST 1992
I'm looking for suggestions on how to blot a sequencing gel. It would be easy
if I could get ahold of one of those neat devices that run the DNA straight off
the end of the gel and onto the membrane, but .....
The details that I'm after include:
- Membrane (nylon or nitrocellulose? (my guess is nylon)
- Transfer buffer? (with nylon its less critical?)
- The basic mechanics of the process: The minimalist approach seems
1) rinse/soak gel in 10x SSC for 20min
2) pick the gel up on 3mm paper, place it on "resevoir" (3x3mm w/SSC)
3) add membrane and "wick" (more 3mm then paper towels).
4) let it go o/n.
if folks out there who have done this before could confirm/correct this.
Nigel Walker \ Mu / (510) 642-7085 Lab
Dept. of Plant Biology \ / (510) 642-4995 Fax
U.C Berkeley \ / (510) 654-LEAF
Berkeley, CA 94720 \/ nigel at notmendel.berkeley.edu
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