QIAGEN native lysis protocol

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Tue Jun 15 13:37:40 EST 1999


Kevin Urbanic <kurbanic at uoguelph.ca> wrote:
> I am currently trying to natively purify a His-tagged protein using 
> QIAGEN Ni2+ resin.  There protocol for native cell lysis is not very 
> efficient.  Does anybody have any real good methods for natively lysing 
> E.coli cells?

I don't know what Qiagen's protocol is and how you have determined that
it is not very efficient, but most lysis protocols use physical methods
for the disruption of the membrane. For example, ultrasound is very
efficient in lysing bacteria, although unsuitable for larger volumes.
Another somewhat more gentle method is using a French press. Certain
bacterial strains (BL21 with pLysS plasmid) can be lysed by simple
freeze-thawing cycles.

HTH,
--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */



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