Transgenic plants : quick discrimination of independent lines ?

Dr S T May lsrei at csv.warwick.ac.uk
Mon Jan 27 13:55:12 EST 1997


Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Transgenic plants : quick discrimination of independent lines ?

PCR will say WHETHER a plant is transformed, as will Ab resistance,
what I think the original enquirer is asking is to distinguish
between different insertions in separate plants
i.e. are the plants descended from a single insertion event.

Correct?

Apart from southerns, the only methods that immediately come
to mind are:

i) digestion, self-ligation and IPCR - definitely more work
than a Southern.

ii) RAPDs with a single LB out primer at consistent but low 
temperature - as long as the transformed population is
inbred (or not too het), this should produce different,
but consistent banding patterns for each insertion event.
One of the bands will be biased by the LB primer to include 
the flanking region.


I hope this helps,

I'd make sure your PCRs are consistant first though ;)

If you require more info or LB out primers (or RB out)
please contact me. I'm working on something that uses
a different aspect of this kind of effect.


Sean.


-- 
-------------------------------------------------
Sean May  [sean.may at warwick.ac.uk]
Warwick University, Coventry, U.K.
http://www.oikos.warwick.ac.uk/~bioys/



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