By s.seekamp on Tuesday, January 14, 2003 - 05:08 am:
Hello everybody!
I am looking for some one who has experience in doing HPLC analysis of olignonulceotides and/or in the quality assurance of amidites.
How do the producers get the coupling efficiencies for there certificate?
Are the doing a homo-coupling or a sequenz?
Do the trust in the numbers of a conductivity measuremnt cell in the oligo-syntheziser or are doing an HPLC analysis?
By Jim Thayer on Tuesday, January 14, 2003 - 05:42 pm:
Regarding coupling efficiencies of oligonucleotides and their evaluation, Tony Yeung has posted a rather elegant (ABRF, Nucleic Acid Research Group) poster at the following link:
http://web-apps.fccc.edu/fccc/narg/
Go to this site, click on NARG 2001 study, then on presentation and poster, then on QC presentation. The poster is downloaded as a PDF file.
The work presented shows several chromatographic approaches to evaluation of coupling efficiency, including IP-RPLC, CE and anion-exchange approaches at neutral and high pH. Dionex markets an anion-exchange column (DNA Pac PA100) that would perform the anion-exchange separations at both pH values shown in the poster.
I hope this helps
Jim Thayer