purifying short probes

upyers mc at u.washington.edu
Tue Feb 27 12:49:45 EST 1996


In article <e.pietrzak.6.313284B7 at mailbox.uq.oz.au>,
e.pietrzak at mailbox.uq.oz.au (Eva Pietrzak) wrote:

>    Hi, netters,
> 
>    Do you have any tips on how to purify a very short 
>    oligonucleotide probe (20 mer) after 33P labelling?
>    Some home-made equivalent of a mini spin-column
>    or a brand name popular enough to be available
>    Down Under?
> 
>    thanks, Eva

There are undoubtably several companies that make such columns (try Qiagen).

However, for a homemade solution;

   Run your probe out on an acrylamide gel (8%). TBE buffer is best.
   Detect the probe either by geiger counter or (better) by shadow casting;
      Hold a UV source near gel. Probe should appear as a shadow on a
      white background.
   Cut out probe band and mince the gel slice.                                
   Put minced slices in 3M NaOAc (pH 4.8) and mix overnight.
   Next Morning spin down gel slices, remove SN (wash gel slices 1x with 3M 
      NaOAc).
   ppt in EtOH.

Good luck

-- 
"The two most common things in the universe are hydrogen and stupidity" Harlan Ellison



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