screening yeast by PCR
FATIMA at AT.AC.UNA.AIMP
Tue Oct 5 04:10:00 EST 1993
Dear Michael and others,
I do not have any real personal experience with PCR screening of yeast colonies
(except of one failed attempt). However, I found the following method in our
lab files and I remember that it worked in other people's hands. I think you
might find it useful. Good luck! Fatima
Really Fast Yeast DNA/Plasmid Prep
by Josh Bonkovsky, Nasmyth lab (modified from Beverly Errede)
Harvest 2 mL overnight culture with 2' 14 K centrifugation. Suspend pellet in
300 5L TE, add 200 5L glass beads, and vortex 15'. Extract with 300 5L phenol,
and then ethanol precipitate, and suspend DNA in 60 5L TE.
For PCR: a 1:10 dilution of the DNA is usually good.
To recover plasmids: sometimes this method is ok, sometimes not. Usually one
should try both undiluted and a 1:10 dilution for transforming bacteria.
More information about the Yeast