electroporation of BL21(DE3)pLysS

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Mon Jul 3 14:00:02 EST 1995


 In article <CDMENU.13.2FF79AA3 at ccs1.cc.monash.edu.au| 
 CDMENU at ccs1.cc.monash.edu.au writes:

| Has anyone tried to electroporate the E.coli strain BL21(DE3)pLysS? If so, 
| could you please send me the protocol for the preparartion of the cells and 
| also the protocol for electroporation.
| My Email address is:  jackiec at ccs1.cc.monash.edu.au
| Thanks very much!
| Jackie 

The problem is that BL21(DE3) is a very poor recipient for any kind of
transformation. The trick is to make your constructs within a different
strain like HB101 or DH5a, confirm the DNA is correct, then re-transform
purified DNA into BL21(DE3)pLysS for expression. You might look at the
original references for the best conditions of electro-transformation of
E. coli cited in this article:

@article{Hengen1995Juntibs,
author = "P. N. Hengen",
title = "Methods and reagents - Electro-transformation of
{{\em Escherichia coli}} with plasmid {DNA}",
journal = "Trends in Biochemical Sciences",
volume = "20",
number = "6",
pages = "248-249",
month = "jun",
year = "1995"}

*******************************************************************************
* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - - - - - Methods FAQ list -> http://www-lmmb.ncifcrf.gov/~pnh/ - - - - - - *
* - - -  Anonymous FTP from ftp.ncifcrf.gov as file pub/methods/FAQlist - - - *
*******************************************************************************



More information about the Methods mailing list