Unbreakable bugs!!!

Paul N Hengen pnh at kaylor.ncifcrf.gov
Mon Jun 17 17:29:14 EST 1996


Philippe Lefebvre (plefebvre at pop.univ-lille2.fr) wrote:

> We have a strange problem here: for two weeks, we have been unable to obtain a
> good and efficient lysis of E. coli strains (for protein extraction purposes).
> We are usin Tris Glucose buffer, to which we are adding lysozyme.  Lysis occurs
> within minutes, usually... We have tried new lyzozyme, pure water, freshly
> transformed bugs, checked pH...  still no lysis.  Does anyone ever encountered
> such a problem?

Try pre-washing the cells with 1M NaCl as in Paul's "coli-blaster"
technique. You can find the whole protocol for plasmid extraction
in the archives of this group.

--
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