oligo purification on hplc

David J. Perry dperry at miu.edu
Sat Nov 5 17:43:16 EST 1994


Are you trying to purify an oligo directly after deprotection or one
that has been partially purified in some way?  We generally purify our
oligos with the trityl group still on by reverse-phase HPLC.  The
trityl group makes the oligo hydrophobic relative to the failure
products; hence the desired product comes off the column well after the
failure products.  Our standard protocol is to inject our oligos on a
Hamilton PRP-1 column and run a gradient of 5-60% acetonitrile in 0.1M
triethylamine acetate (pH7.0) over a period of about 8 min.  We like
the PRP-1 column because its polymeric support is stable at high pH so
we can inject our samples in the ammonium hydroxide they were
deprotected in.  

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.

Best of luck!



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