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.
> 
> 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*.

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

 
> Regards
> Bob DeLisio

Bob,

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.
  
-- 
Cheers,

Martin

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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