[Protein-analysis] Osmotic shock - solvent vs culture volume?

Julio juliocortazar at o2.pl
Sun Mar 19 10:20:43 EST 2006


Hi everyone.

Have a question about standard procedure of osmotic shock. I grow my
E.coli after induction by means of IPTG for 12-16h in 27Cdeg. Than I
want to make osmotic shock. Many people say many things about
proportions of Tris, sucrose, EDTA solution vs volume of culture after
harvesting by cetrifugation. For me the lower volumes  the better
situation for next steps (eg batch with Ni-NTA resign). I read about
80mL of Tris, sucrose, EDTA solution for 1g of culture after spin but
also 8mL of this solution for 100mL  of culture volume befor spin. What
do you advise me?

JC



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