How to extract DNA of Bacillus subtilis efficiently? - (Nov/27/2005 )
Could you tell me how to extract DNA of Bacillus subtilis efficiently?Please give me some advise.Thank you!
Chromosomal DNA, plasmid DNA, or genomic DNA?
genomic DNA,thank you!
Well, when I was working with Bacillus anthracis, we recovered genomic DNA by using an alkaline lysis method that included heating the lysis step for 10 minutes at 65°C, and recovering the DNA by ethanol precipitation from the cleared lysate.
This was twenty years ago, and there may be better methods now, but this worked well for us then and was quite easy.
You could also try resuspending the cells in TE/SDS/Proteinase K, incubating at 56°C for an hour or so, extracting with phenol, then phenol:chloroform, then with chloroform until there's no interface, and ethanol precipitate your DNA from that.
I'll try this method recently,by the way ,did you prepare the genomic DNA for the templet of PCR?Thank you !
When I worked with B. anthracis, PCR wasn't invented yet...
But, we use similar procedures (TE/SDS/PK) to get genomic DNA from Gram negatives, and it works fine for PCR...
Oh, ,right,thank you
Could you gei me something detail about the procedures (TE/SDS/PK)?Thank you!