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E. coli cell lysis for Protein Isolation URGENT HELP PLEASE

Rafael Maldonado rmaldonado at ua.es
Fri Jan 14 12:51:21 EST 2000



Dr Mick Jones wrote:
> CAN ANYBODY HELP?
> 
> What I need is a simple method for lysing the cells that doesn't use a
> sonicator.

You may treat the cells with lysozyme (0.5 mg/ml) for a couple of hours.
Then freeze the cells and de-freze several times. Some DNAase could be
useful to advoid excessive viscosity due to chromosomal DNA. This is a
protocol from Pharmacia, used for GST protein extraction, so must be OK.
E-mail me if you cannot find it in the Pharmacia webpage.

-Rafa

-- 
Rafael Maldonado
Divison of Genetics
University of Alicante (Spain)




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