Linear Acrylamide Carrier

George W Chacko gchacko at magnus.acs.ohio-state.edu
Thu Dec 19 15:43:40 EST 1991


This is actually a follow up to the glycogen query. I got so many 
messages that I thought I would just post the reference.

Cell 37:889-901 (1984)
F Straus and A. Varshavsky.

5% acrylamide (no bis) in 5mM TRis, pH 7.4 + 0.1 mM EDTA
de gas
ammononium persulfate to 10 mg/ml
TEMED 75 ul per 100 ml

Let it sit until it becomes viscous (it will not form a gel in the absence of 
a cross linking agent) --hence linear acrylamide.

Stocks:

40% acrylamide
25% AMPS
TEMED
1M Tris-HCl pH 7.4
0/.5 M EDTA pH 8.0

Note: *I* (so that you're aware of the uncertainty involved) have used just
5% acrylamide without polymerising it *once* and the precipitation came
off OK.

I use 1ul linear acrylamide + 1/10 vol of 3M NaOAc pH 4.2 + 2 vols
of absolute ethanol for anything upto 50 ul and proportionately  thereafter.
You can see the precipitation immediately but I popo into the -20 for
10 minutes after that and spin.

I haven't tried it for RNA.


George



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