help: amplification,polysaccharides (complete version)

monnerot at cgmvax.cgm.cnrs-gif.fr monnerot at cgmvax.cgm.cnrs-gif.fr
Tue May 9 11:31:42 EST 1995


To everybody, concerning problems of amplification of microsatellites

I am working on a sedentary polychaete (P.koreni) and have already isolated
and sequenced microsatellites of the type (TC)n, (TG)n and (AT)n.
I designed primers for some loci, but I am not able to amplify any of those
using genomic DNA. The primers seem to be o.k. as the minipreparation of 
the plasmid containing the respective locus amplifies without any problems.
So I feel that my problems are due to some inhibitor in my DNA extractions, 
perhaps polysaccharides. Has anybody got any idea how to solve this problem
?I already tried the standard CTAB-extraction but my DNA is still 
unrestrictable and unamplifiable. I would be grateful for suggestions as to 
how to possibly modify this protocol or for any other protocol likely to 
separate polysaccharides from DNA. Thank you all very much for your 
attention, yours sincerely
				Gudrun Weinmayr

My E-mail is weinmayr at sb-roscoff.fr; Faxnumber: (33) 1 69 07 04 21




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