Bacteria cell lysis

Andy Kravetz akravetz at MO.NET
Mon Feb 27 13:44:19 EST 1995


Brian Carey (bcarey at zookeeper.zoo.uga.edu) wrote:
: Dear Netter, I discovered tonight that I can't lyse bactera. Would
: appreciate all suggestions. Here's my story. The cells are BL21(DE3).
: Cells from 200 ml culture OD600=1.0 was suspended in 5 ml of high salt
: lysis buffer (no lysozyme), freezed/thawed 2x before and 2X after 10
: cycles of 30 sec sonication at max setting. The pellet I got after spining
: was full of unlysed cells. I thought maybe the whole mass was too
: concentrated. So I resuspended the cells again, added another 5 ml of
: buffer, sonicated 20 X 30sec and spin. I still got a huge pellet of
: unlysed bacteria cells. I'm trying to purify over-expressed protein in
: these bugs and would like to avoid the use of lysozyme if possible.
: Any idea?
: Thanks in advance.

: Tari
: University of Georgia 
: Cellular Biology Dept.
: E-mail: wll at zookeeper.zoo.uga.edu

--

Have you tried NaOH/SDS as a lysis buffer??


------------------------------------
//"To leave the world a bit better place, whether by a healthy child, a //
//garden patch, or a redeemed social condition; to know that even one life// 
//has breathed easier because you have lived. This is to have succeeded."//

Andy Kravetz
Freelance Journalist
St. Louis, Missouri
akravetz at mo.net



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