total proteins from B.subtilis

Michael Witty mw132 at mole.bio.cam.ac.uk
Sun Aug 19 07:58:22 EST 2001


Dear Jayakumar,
              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
                                50microlitres 10%SDS
                                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:

> HI..
>     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
> Sincerely
> Jayakumar
>
>
> ---
>




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