stop growing of E.coli
ekhatipo at NOSPAMmidway.uchicago.edu
Mon Jan 22 14:51:05 EST 2001
I am almost sure that nobody did that, but here is my suggestion. Poor
acetone into your culture up to ~40% v/v (should try different
concentrations for optimal results), mix, keep the mixture in the cold room
for a couple of hours to allow precipitation, spin down precipitate,
resuspend the pellet in the working buffer.
Another suggestion, use mild detergents like CTAB to permeabilize cells.
Cells can be spun down after ~1-2-min incubation with CTAB (~0.1 mg /ml -
best concentration should be determined in advance). However, you most
probably will have to use a more harsh methods of extraction like
French-press or grinding in a mortar with sand - cannot use sonication
efficiently since cells are permeabilized and thus would not resonate.
Third method - use toluene for permeabilization. But there is no guarantee
that, as with CTAB, nothing will happen with your protein of interest (like
proteolytic degradation) during subsequent handling.
Hope this helps,
"Shang Hao" <shangh at ecn.purdue.edu> wrote in message
news:3A68D2B2.56A6B07F at ecn.purdue.edu...
> I want to stop growing of E.coli in expotential phase quickly and
> extract proteins for E.coli. Could you give me some suggestions about
> it? Your help is gratefully appreciated!
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