help lysing E. coli to purify inclusion bodies

[jfh]

Howdy all, 

I (we) are making mamalian proteins in ecoli using pT7 and BL21(DE3).

I've had success with the system but my lysis is always a crap shoot. 
Sometimes lysis occurs easily sometimes not.  I have been following the
methods in enzymology, nagai and thorsen protocol, using lysozyme,
followed by deoxycholic, NP40, and DNAse 1.

What do other people do?  Post some suggestions here or email me.


John Hess, PhD                    Phone me 916 752 8420
Dept of Human Anatomy             FAX me 916 752 8520
University of Calif               Email me jfhess at ucdavis.edu
Davis, CA                         or leave me alone, your choice.