G-C content

[Andreas Brune]

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. 
Bacteriol. 39:159-167

>Does
>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).

   DSM
   Mascheroder Weg 1b
   38124 Braunschweig
   Germany
   
Regards,

Andreas



Dr. Andreas Brune         	Phone: 	+49-7531-883282
Mikrobielle Oekologie     	Fax:   	+49-7531-882966
Universitaet Konstanz     	E-mail:	Andreas.Brune at uni-konstanz.de