Q: Bacillus subtilis genome DNA extraction protocol?
W. C. Lee wrote:
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
Re: Q: Bacillus subtilis genome DNA extraction protocol?
I am isolating genomic DNA from Bacillus subtilis using the prescribed protocol. But when i am running the genomic DNA in 0.8% agarose the genomic Dna is not running through the gel, it is stuck to the mouth of the gel. Please suggest some quality control for the process.