mkennedy at chmeds.ac.nz
Sat Apr 24 03:25:35 EST 1993
In article <9304222010.AA09552 at mailgate.roche.com>, delisior at rnisd0.DNET.roche.com (Bob DeLisio) writes:
> Hi Pat:
> I am posting this on the net because I think others might find it useful.
> PCR COLONY MICROSCREEN
> 1. Pick colonies into 100 microliters of LB+antibiotic in a sterile
> microtiter well.
> 2. Grow 2-4 hours at 37 C on shaker platform until turbid.
> 3. Add 2 microliters of the culture to 23 microliters of PCR reaction mix*.
> Bob DeLisio
This looks fine, but you could save yourself a couple of hours (and some
L-broth)! by transferring the colonies into 0.5ml of water and doing the
PCR directly on that. See Gussow and Clackson, 1989, NAR 17, p4000; they
recommended boiling the cells first, but it's not necessary; you can rescue
viable cells from after transferring them into water.
NNNN NN Martin A Kennedy (E-mail = mkennedy at chmeds.ac.nz) ZZZZZZZ
NN NN NN Cytogenetic and Molecular Oncology Unit ZZZ
NN NN NN Christchurch School of Medicine ZZZ
NN NNNN Christchurch, New Zealand ZZZZZZZ
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