Q: Bacillus subtilis genome DNA extraction protocol?

alejandra.martinez at biotek.lu.se alejandra.martinez at biotek.lu.se
Mon Jul 7 03:57:30 EST 2003


W. C. Lee wrote:

> Dear b.m.methds-reagnts readers,

> Do anyone know the easy and quick protocol to
> extract genomic DNA from Bacillus subtilis?
> I used a plant genomic DNA extraction kit from
> Qiagen, which was the only kit available to me,
> and couldn't get my PCR done.

> Thanks in advance,

> Woo

I routinerly use this method with excellent results in several Bacillsu species:
To obtain genomic DNA, 1.5 ml of each overnight culture
was centrifuged at 13,000 g for 3 min. The pellets were resus-pended
in 1 ml of 10 mM Tris HCl, 10 mM EDTA, 100 mM
NaCl, 2% (v/v) SDS, and 400 mgml –1 proteinase K; the mix-ture
was incubated at 56°C for 30 min. After phenolization
and chloroform purification steps, DNA was precipitated
with 1 volume of 2-propanol, washed twice with 70% (v/v)
ethanol, and finally resuspended in 200 ml of sterile water.
I hope it hepls



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