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Problems in CaCl2 method ofr competent cells - (Nov/08/2007 )

Hello everybody

I'm having problems in making competent E. coli cells. I use the CaCl2 method, but my cells do not transform correctly after the heat-shock treatment. I´ve alredy tried with both BL21DE3 and BL21DE3 STAR cells, but they do not go on!!!

Am I going wrong in some part of the method? What could cause this? Does anybody knows???




change your competent bacteria stocks or diminish Atb concentration