Why are my electrocompetent bugs lysing?

chenlinan at 263.net chenlinan at 263.net
Thu Jan 20 08:29:04 EST 2000


I met the same problem several times. Tomorrow, I will repeat once. I think centrifuge and resuspend force should be reduced, which lowers the possibility of cell lysing. When dissociating tissue in primary cell culture, I met the similar problem, and sovle it using the same methods.

Any other suggestion?

Chen Linan
Dept. of Anatomy
Beijing Medical University

-------------
>Help! I have been making electrocompetent bacteria for years and for some
>reason my bugs are lysing after the first wash. By the second wash there are
>very few cells left. I have tried both XLI-BLUE and TOP10, different media,
>very cold water, not quite so cold water, faster wash times and more gentle
>washing with no success. What am I doing wrong?
>
>Rebecca
>________________________________
>
>Rebecca Ormsby.
>Dept. Microbiology & Infectious Diseases,
>Flinders University of South Australia
>Bedford Park, S.A. 5042
>
>phone: (+61) (8) 8204 5252       ___o
>                                                      _ \    <_
>                                                    ( *)/     (* )
> fax: (+61) (8) 8276 8658
>
>---

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