Bacteria cell lysis

Cornelius Krasel krasel at alf.biochem.mpg.de
Sun Feb 26 12:08:27 EST 1995


Brian Carey (bcarey at zookeeper.zoo.uga.edu) wrote:

[he or Tari cannot lyse BL21(DE3) by 10 x 30 sec sonication]

You could try the usual stuff like detergent, french press, ... However,
if you want to stick with sonication, just do low-speed centrifugation,
resuspend the pellet and sonicate it again. Repeat this two or three
times.

Efficiency of sonication is also dependent on the size of the tip. It
is almost senseless to sonicate a 10 ml probe with a microtip. If you
can attach a macrotip to your sonicator, do it. I sonicate 7.5 ml
bacterial suspension from 1 l culture for 2 x 10 min with a macrotip
(at most 20% max. intensity, 50% duty cycle) and find almost only
outer membrane in the remaining pellet. Using a microtip gives much
less cell lysis.

Hope that helps,
--Cornelius.

--
/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany  */
/* email: krasel at alf.biochem.mpg.de                 fax: +49 89 8578 3795  */
/* "Science is the game you play with God to find out what His rules are." */



More information about the Methods mailing list