Isolation of "hot" oligos

Suzy Paalman suzy at welchlink.welch.jhu.edu
Sat Jun 15 19:49:43 EST 1996


In article <kremers-1306961122030001 at ifepmac3.uni-muenster.de>,
kremers at uni-muenster.de (Joachim Kremerskothen) wrote:

> Hi netters,
> 
> we have a problem with the isolation of radioactive-labeled oligos
> (20mers) after T4 kinase phosphorylation. Ethanol precipitation and gel
> filtration with Pharmacia`s G50 columns didnt work very well (maybe
> because of the lenght of the oligos?). Are there other and better methods
> for the isolation?
> 
> -- 
> Dr. Joachim Kremerskothen
> Institut for experimental Pathology
> University of Muenster
> Mendelstrasse 11
> D-48149 Muenster
> Germany
> phone ++49 251 838615
> fax ++49 251 838616

Hi, 

I routinely label and purify 20mers.  The two methods I use to get rid of
free label are G-25 columns (as someone else suggested, G-50s won't work
for 20mers) and ethanol precipitation with 10mM MgCl2 as the salt (if
possible-I use sodium acetate if I've inactivated the kinase with EDTA) and
using linear polyacrilimide (available from Sigma) as the carrier.  Both of
these methods work well for me.

Suzy

-- 
Johns Hopkins School of Medicine



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