Oligonucleotide analysis by anion exchange
Posted: Fri Jan 18, 2008 3:01 pm
by grrr_aham
I would appreciate any advice or comments on the Dionex DNAPac PA200 hplc column for the separation of oligos. We need maximum resolution. It seems to be the most advanced anion exchange column on the market. Is it as good as the makers claim, or could anyone recommend an alternative? Are these columns as robust as a modern C18 5uM column?
Thanks!
Posted: Mon Jan 21, 2008 5:35 am
by Kelly Johnson
I am not familar with the Dionex product performance (or its stability) but ZirChrom does have a robust anion exchanger that may interest you. Oligonucleotide application notes can be found on our website at:
http://www.zirchrom.com/pdf/267.pdf
http://www.zirchrom.com/pdf/166.pdf
Hope this helps!
Kelly
Posted: Mon Jan 21, 2008 7:03 am
by XL
grrr_aham,
There are alternatives for this application, such as by RPLC. However, when it comes to anion-exchange method, Dionex DNAPac PA200 is the most advanced anion exchange column on the market and does perform as claimed in terms of resolution, ruggedness, reproducity, and column life time. Here is a link where you will find more information:
http://www1.dionex.com/en-us/columns_ac ... 60685.html
In case you have specific concerns or questions, please feel free to ask.
Posted: Mon Jan 21, 2008 9:41 pm
by Jthayer
Dear Grrr_aham,
My name is Jim Thayer, and I was part of both DNAPac column development teams. We are very proud of our accomplishments with these columns, and often communicate with customers with specific recommendations to specific questions.
I am happy to offer to help you with your application. In addition to elution of oligos in order of their length (as supported also by IP-RPLC) the capabilities offered by the DNAPac columns included:
1) resolution of n, from n-1 oligomers for normal length oligonucleotides (with examples up to 75 bases);
2) Control of selectivity for very similar oligos by modulation of eluent pH, salt-form, and solvent concentration;
3) Resolution of unreacted oligos from those derivatized with virtually all of the popular fluorophores used for capture and reporting probes;
4) Control of poly-G tract hydrogen bonding by high pH chromatography; 5) separation of many related morpholino-linked oligonucleotides;
6) resolution of oligonucleotides with phosphoramidate linkages from their normally-linked counterparts;
7) resolution of fully phosphorothioated oligos from those harboring one, or more, residual diester linkages; and
8) Resolution of RNA oligomers harboring one, or more, aberant 2´-5´ internucleotide linkages from those having only normal 3´-5´linkages.
This list of capabilities includes only those that have been publicly reported to us, or which were developed in our labs in Sunnyvale.
If you wish to discuss your specific application with us please contact me through Dionex Corporation (408) 737-0700, and ask for Jim Thayer.
Regards,
Jim