Cell disruption without sonication

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Mon Jul 26 09:48:24 EST 2004


voyager wrote:

> Help !!!!
> 
> I am expressing my protein in E. coli BL21, and I am looking for
> another way to disrupt the cells without sonication
 Does anyone have
> a protocol ?!?!?!  I tried lyzosyme followed by frezze-defrezze
> methodology, but the separation of pellet, supernatant and debrit was
> terrible
 I needed to centrifugate several times, so I am not sure
> about the integrity of my protein
 I am growing a culture of 50 mL.

You can use a French Press, with or without pre-treatment with lysozyme.
In a French press the sample is passed under high pressure through a
narrow orifice, the resulting shearing forces disrupt cells, yeasts and
bacteria.  



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