Tritech_Research at lamg.com
Sun Dec 1 16:52:52 EST 1996
S L Forsburg forsburg at salk.edu writes:
Plasmids from yeast lysates are notoriously hard to transform
into E. coli. Probably some inhibitors come along. However,
we have increased our efficiency by several logs by using
ELECTROPORATION to transform our bugs. It really makes a
huge difference, at least in our hands (yes, from both
pombe and cerevisiae lysates).
So, assuming you saved your lysates....prep up some of your
HB101 to make them electrocompetent, and zap them. You
won't get a lawn of colonies but you will get enough to work on.
Also, remember that yeast, unlike bugs, can be multiply transformed
with different plasmids containing the same replicon. Your
transformants may carry more than one plasmid, so bear this in mind.
We make a very cool hand-held cordless rechargeable electroporator,
called The Cloning Gun, which is pre-optimized for exactly that kind
of transformation. It's only $995, and uses pipetting electrode
assemblies (called Pipectrodes) instead of cuvettes, and they are
only $1.50 each. If anyone would like more info, and/or a copy
of our catalog, please e-mail, or call me at 800-7-333-874.
I apoligize if this is too "spammy" - we designed it to help by saving
labs time and money....
Back to the sidelines,
TRITECH_RESEARCH at LAMG.COM
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