PCR leaf

Malcolm M. Campbell malcolm.campbell at plants.ox.ac.uk
Thu May 15 08:50:54 EST 1997


Hi Yvonne,

In reply to your querry about quick and easy ways to prepare leaf tissue
"template" for PCR reactions, check out:

Alkali treatment for rapid preparation of plant material for reliable PCR
analysis. Plant J., March 1993, Vol. 3, p. 493

It's a quick, easy and relatively reliable (given how "quick and
dirty" it is) protocol from Jonathan Jones' lab.  We've used it a couple of
times to verify the presence of the transgene in large numbers of
Arabidopsis transgenic lines.  We've not had a problem with false
positives.  False negatives, on the other hand, have been a problem. 
We're sticking with DNA preparation a la Dellaporta for this reason.  The
Dellaporta protocol is not as
simple, nor as rapid, as the alkali prep, but the time investment is worth it
if one wants to ensure that all putatively transgenic lines are verified
by the screen.  Presumably the same consideration would be pertinent if
one was introgressing a gene as well.

I hope this is useful.

I trust all is well in Waterloo.  Please say hello to Barb Moffat for me.

BTW, there is something not quite right with your return address such that
any attempt at a direct reply gets bounced by the watstar server.

All the best,

Malcolm

-- 
Malcolm Campbell                 malcolm.campbell at plants.ox.ac.uk
Fibre Biotechnology Lab          44.1865.275135 (voice)
Department of Plant Sciences     44.1865.275074 (FAX)
University of Oxford
Oxford OX1 3RB, UK



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