PAGE of short DNA fragments

chmc chmc at chmc.org
Thu Mar 16 12:22:21 EST 2000


I have only done this a couple of times, but what I did was:

Made 30% acrylamide solution:
29g acrylamide
1g N,N'-methylenebisacrylamide
H2O to 100 mLs
Filter

The mini gels that I used required 5 mls of solution, and I ran 12% gels.  To do
this, I added 2 mLs of 30% acrylamide, 1.95 mLs of H20, 1 mL of 5X TBE, and 0.05
mLs of 10% Ammonium persulfate.  Add 2 microliters of TEMED, pour the gel, and
let it polymerize.  Put the gel in the tank with 1x TBE, and wash the wells out
with a syringe or pasteur pipet.  Load the samples with regular old DNA loading
dye, and run it.

Adam



Keith O'Neill wrote:

> Hi,
>
> Does anyone know of a basic protocol or reference for running non-denaturing
> polyacrylamide gels for short double-stranded DNA fragments.
>
> Thanks in advance,
>
> Keith





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