oligo purification on hplc

Lawrence Washington lwashing at sunflower.bio.indiana.edu
Wed Nov 9 15:47:07 EST 1994


> If your query is about oligos whose trityl group has already been
> removed, the standard HPLC technique seems to be anion exchange
> chromotography. We, unfortunately, have no experience with that.
 
	We have been ion exchange purifying our oligos for several years, and it
works quite well.  The buffers are simple and safe (relative to
acetonitrile) but it does not work as well on long (30+ bases) oligos and
the oligos come off the column in high salt.  We use a "Mono Q" anion
exchange column (Pharmacia).  Buffer A is 10 mM NaOH.  Buffer B is 10mM
NaOH, 1.5M NaCl (what could be simpler?).
	The full length oligo comes off in about .65 M NaCl and we usually desalt
it before using. 


Lawrence Washington
Indiana University
Institute for Molecular
 and Cellular Biology



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