Andreas.Brune at uni-konstanz.de
Fri Mar 10 15:49:35 EST 1995
In article <paul_b-030395085101 at clone2.mcb.uconn.edu>,
paul_b at biotek.mcb.uconn.edu (paul betts) wrote:
>We have recently tried a protocol designed for determining genomic DNA base
>composition. The protocol requires hydrolyzing DNA in formic acid then
>resolving the constituent bases using HPLC with a "Partisil" strong anion
>exchange column with an isocratic elution profile. This method did not
>work in our hands, though admittedly the SAX column we used was not a
>Partisil. We tried several permutations of this protocol but failed to
>resolve the constituent peaks. Does anyone out there have an alternative
>protocol for HPLC for this purpose that they are willing to share?
We are successfully using the following method which degrades the DNA enzymatically
and separates the nucleosides by reversed-phase HPLC.
Mesbah M, Premachandran U, Whitman WB (1989) Precise measurement of the G+C content
of deoxyribonucleic acid by high-performance liquid chromatography Intern. J. Syst.
>anyone know of a facility that performs base composition analysis (HPLC or
>other) as a service? We have a new strain of Mycoplasma that we are
>characterizing and wish to know its G-C content.
Try the German Collection of Microorganisms (DSM).
They peform G+C content analysis as a service (reasonable fee).
Mascheroder Weg 1b
Dr. Andreas Brune Phone: +49-7531-883282
Mikrobielle Oekologie Fax: +49-7531-882966
Universitaet Konstanz E-mail: Andreas.Brune at uni-konstanz.de
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