total proteins from B.subtilis
mw132 at mole.bio.cam.ac.uk
Sun Aug 19 07:58:22 EST 2001
to make total proteins form E. coli I do this:
1]. Spin down 1ml of E. coli overnight culture.
2]. Add 200microlitres thus: 50microlitres H2O and resuspend cells
100microlitres 2x loading buffer and mix
3]. 100 degrees C for 3 minutes (time depends on individual protein).
4]. Load 15microlitres on a minigel for Coomassie staining
Sometimes it is a bit viscose and I pass the extract through a needle 5x
to shear DNA.
Maybe this would work for B. subtilis too. Regards, Mike.
On 19 Aug 2001, R. Jayakumar wrote:
> I need a simple and quick procedure for extracting total proteins from
> vegetative and sporulated cultures of B. subtilis for doing an SDS-PAGE.
> Any help would be very much appreciated.
> thank you
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