Hi
We occasionally use a Waters DEAE-5PW column for preparative oligo
purification and use sodium perchlorate (lithium perchlorate can also be
used) in neutral (unbuffered) solutions. We also use a column oven and
run at 50 degrees. This seems to work reasonable well and might be the
only way to denature oligos when high pH is not an option. If you're
interested in a reverse-phase trityl-on HPLC method using NaOH, you can
see M. W. Germann et. al., 1987, "A general method for the purification
of synthetic oligodeoxyribonucleotides containing strong secondary
structure by reveresed-phase high-performance liquid chromatography on
PRP-1 resin", Analytical Biochemistry 165, 399-405.
Alain LAURENT wrote:
>
> Hi All's
>
> I am using a WATERS GEN PACK FAX Column (anion exchange HPLC) to analyse
> oligonucleotides. It works very well but it seems that I cannot use NaOH
> to increase the pH and denaturate the oligos. What kind of basic buffer
> could I use instead ? (as easy to prepare and to store as NaOH )
>
> Any people used to work with this column I could talk with ?
>
> Thanks for your comments
>
> Best regards
>
> Alain
>
> **************************************
> Alain LAURENT Ph. D
> DNA&Peptide Chemistry
> ESGS Groupe CYBERGENE
> 11, rue Claude Bernard
> 35 400 Saint MAL0
> FRANCE
> Tel: +33 (0)2 99 21 90 40
> Fax: +33 (0)2 99 21 90 41
> e-mail: a.laurent@eurosequence.com
> http://www.eurosequence.com
> **************************************
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