Ethanol precipitation w/ linear polyacrylamide

Benoit_Hebert at IAF.UQUEBEC.CA Benoit_Hebert at IAF.UQUEBEC.CA
Thu May 26 22:36:54 EST 1994


Here is the reference I promised:

Nucleic Acids Research  18(2): 378   (1990)
by
Claire Gaillard and Francois Strauss


I have tried this with precipitation RNA, cDNA, DNA and it does wonders!

The sequencing service at the Institute now recommends this protocol.

Outline of linear polyacrylamide solution preparation:

prepare a fresh 5% acrylamide sol. (WITHOUT BIS !) in 40mM Tris-HCl, 20mM 
NaOAc, 1mM EDTA, pH 7.8. Add 1/100vol of 10% APS and 1/100 vol TEMED.
Let pol. for 30min. PPtate the viscous soln w/ 2.5 vol EtOH, spin and 
redissolve the pellet in 20 vol. distilled/sterile H2O o/n by shaking.
The 0.25% linear acrylamide can be stored at 4C for years.

We have been using the same for a little over two years w/o any changes...


Benoit Hebert

========================================
Benoit_Hebert at iaf.uquebec.ca

Armand-Frappier Institute
Virology Research Center
Laval, Quebec, Canada    H7N 4Z3
phone: (514) 687-5010, ext. 4636
fax: (514) 686-5626
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