Lysis of Listeria and other gram positives

BehariRN beharirn at CCMAIL.APLDBIO.COM
Wed Sep 27 10:27:24 EST 1995


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Hello folks,


I am trying  to get quantitative lysis of Listeria and other gram poitives in 
comples food matices and having problems. I want a quick method that is not 
labor intensive and there are too many samples to process in a day. I have tried 
mehtods like Guanidinium and other chaiotropic agents including detergents 
(Triton, SDS); I have also tried using lysozyme and heat shocking the cells 
prior to lysis. The recovery is not more than 10% based on a rough estimate of 
the final concentration of the DNA obtained. I have done these calculations 
based on the genome size of E.coli because I don't know what the genome size of 
Listeria is; does anybody else know? Does anybody else have any other good ideas 
on quick, high efficiency, sample preps from gram positives, pretty please?

THANKS FOR YOUR INPUT.

Richa





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