DNA extraction: Lactob.

Heitor Luiz da C. Coutinho heitor at genes.icgeb.trieste.it
Tue Oct 25 08:57:15 EST 1994


Hi netters,

I have been having problems for extracting genomic DNA from cultures of 
lactic acid bacteria (Lactobacillus fermentum). I have tried the 
guanidium thiocyanate method, but the outcome has been degraded DNA in 
low yields. A lot of "dna" is formed when ethanol precipitating, but most 
of it remains undissolved in the TE solution at the end. We have tried a 
lot of variations, like overnight lisozyme, previous proteinase K 
digestion, more or less chloroform extractions, more or less time and 
temperature of 
incubation with the guanidium solution, checked pH of all reagents, etc.

Has anybody got a tailor-made protocol for extracting genomic DNA for PCR 
and other molecular works with Lactobacillus fermentum (or relatives)?

Thanks for any help.

Cheers from the tropics,

Heitor L.C. Coutinho
Funda at ao Tropical Andre Tosello
Campinas - SP, Brazil
 



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