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freezing comp. yeast cells

Kathy Shire shirek at FHS.CSU.MCMASTER.CA
Tue Jul 26 16:11:53 EST 1994


Yeast Club Members-

I'm a newcomer to the yeast scene.  The novelty of the bread smell has
worn off a while ago, but I'm still an amateur.  I have tried something
that works pretty well and saves me a lot of time.  Someone else might get
some use out of it.
Freezing Competent Yeast Cells:
Small Scale Transformation Using LiAc (Current Protocols in Molecular Biology)
After the 1hr incubation at 30C in 500uL 0.1M LiAc, freeze cells (as is)
at -70C in 100uL aliquots.  Thaw cells when needed at RT, add plasmid
+hsDNA and continue normally.
I get about 500colonies/ug.  The frequency is low but if you want a
plasmid in your yeast in a hurry, it works every time!

Kath






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