Quick PCR

Martin Kennedy 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.
> 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*.

stuff deleted......................

> Regards
> 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

More information about the Methods mailing list