BL21(DE3) pLYSe/s competent cells

[Rico Laage]

Dr. Duncan Clark wrote:
 

Rico
 
> 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
competence 

Rico


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