BL21(DE3) pLYSe/s competent cells

Rico Laage rico.laage at
Wed Nov 6 04:48:41 EST 1996

Dr. Duncan Clark wrote:

> Hi Folks,
> Has anyone had any hassle making electrocompetent cells of the above.
> Thawing from -70C lyses our supposed electrocompetent stocks. I know
> they carry T7 lysozyme but I wasn't aware they were that sensitive to
> one freeze thaw. I'm now going to make ordinary competent cells and use
> them immediately but would like any comments.
> Duncan
in my opinion it is better to use 7% DMSO than using 20% glycerol I use
the protocol of Inoue et al (but growing the bugs at RT)  gives good


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