Looking for suitable bacterial strain

GIETZ at bldghsc.lan1.umanitoba.ca GIETZ at bldghsc.lan1.umanitoba.ca
Thu Jan 21 15:49:00 EST 1993


%%%%%%%   Mike writes    %%%%%%%%%%%%%%

        Hello all.  I have been using Stratagene's SURE bacteria and XL-1
Blue as well as BRL's DH10B electrocompetent cells.  What I need to use is
a strain that tolerates plasmids with minimal rearrangement (like SURE),
which works well with electroporation and (the kicker here...) which
does not have dam or dcm methylation.  If anyone has used a strain for
electroporation which is dam- and dcm-, I'd appreciate hearing from you.

Mike Coady
COADY at ERE.UMONTREAL.CA
%%%%%%%%%%%%%%%%%%%%%%%%%%%%
Hi Mike
        I have used JM110, which is dam,dcm , in a quick and dirty
electroporation and it worked fairly well.
  Unfortunately, I don't have any numbers for you.
I just scraped off a big blob of cells from a fresh O/N plate and
washed with 1 ml of cold water twice and resuspended in 50 ul
of 10% glycerol and EPed.  I plated 2 ul of the 1050 ul of SOC culture
and got a fair number of amp r colonies, if I remember correctly.  
Unfortunately
It's not as good as DH5 alpha done this way.
The genotype is F' traD36 lacIQ|(lacZ)M15 proab/rpsL (Str r)
    thr leu thi lacY galK galT ara fhuA tsx dam dcm supE44
    |(lac-proAB).

Dan Gietz
U of Manitoba
GIETZ at BLDGHSC.LAN1.UMANITOBA.CA



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