Harry.Witchel at Bristol.ac.Uk
Thu Jan 8 07:55:12 EST 1998
For easy protocols (eg transfection of CHO) I do not think that endotoxin
removal is "necessary". I have transfected plasmids growing in XL1 blue MRF' into
CHO using nothing more than a standard Promega Wizard plus miniprep. However, my
transfection efficiency under those circumstances dropped from 60-70% down to 30%.
I would think that if you were trying to transfect a tricky cell line with a
typically low transfectable efficiency, the loss in efficiency associated with
lipopolysaccharides would be undesirable. Just as a reminder, you can greatly
reduce the LPS in your DNA prep just by washing your bugs once in .5 M NaCl before
starting the rest of your maxi/mini prep.
Wolfgang Schechinger wrote:
> Hi all,
> Since there are now maxi prep columns on the market with an endotoxin
> removal feature, I'm interested in when this is necessary.
> When plasmids are grown in strains that produce a lot of
> lipopolysaccharides (LPS), e.g. DH-5, this really might be necessary,
> but with a low LPS strain as XL1, is this true, too?
> What's your opinion or experience?
> This message is RNAse free - please don't touch!
> Wolfgang Schechinger
> University of Tuebingen
> email: wgschech at med.uni-tuebingen.de
> !Junk mail is *not* appreciated!
> SPAMs will be answered automatically by sending my 128MByte virtual memory file.
> This is *NO* joke.
> usual disclaimers apply
Harry J. Witchel, Ph.D.
Bristol BS8 1TD
Harry.Witchel at Bristol.ac.uk
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