electroporation/use no steril glycerol

gwallmanning at wnmeds.ac.nz gwallmanning at wnmeds.ac.nz
Sun Jun 14 21:13:28 EST 1998


Hi!

Or you can omit this step altogether and just wash your cells in water.  I
have (for a long time) centrifuged and resuspended in around half volume of
water, centrifuged and resuspended in a quarter volume, centrifuged and
resuspended in the dregs of water that remain in the tube.  This works
extremely well if you are gonig to use the cells straight away, and you can
save leftovers in 10% glycerol later! ;-) P.S.	We only filter sterilize
glycerol for these cells.


Glenn


In article <6lsf3s$eeu$1 at news2.isdnet.net>,
  "Nougayrède Jean-Philippe" <jpnoug at hol.fr> wrote:
>
> Hello.
> We currently electroporate E. coli cells with the BioRad apparatus; we
> filter-sterilize (0.22) the 10% glycerol in high quality water... using an
> autoclaved stock of high grade glycerol. The efficiency is very good, even
> with wild type strains. Biorad has gone crazy, or what ? ;-)
>
> JP Nougayrede.
> Mol Microbiology Lab. Toulouse. Fr.
>
> Christian Ebeling wrote in <358158B1.1CB1 at mail.mpiem.gwdg.de>...
> >hi bionetters,
> >anybody knows why BIORAD write in the manual "High Efficiency
> >Elektroporation of E.coli":...Do not autoclave or filter-sterilize the
> >glycerol solution...? I do everything to avoid contaminations, but in
> >this step of the manual I have to work not steril!?
> >ebel
>
>


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