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T-DNA rescue

Sun Jul 28 09:40:00 EST 1991

A number of labs are isolating mutants generated via T-DNA insertions.  We
have recently learned that a number of people are having difficulty in cloning
the T-DNA and junction fragment using plasmid rescue.  We have successfully
isolated a number of T-DNA inserts and junction fragments with plasmid rescue.
Two points seem to be key:  First, make sure you have a high E.coli transforma-
tion method (at least ten to the ninth).  Second, the left border seems to be
easier to clone than the right.  This means you will get internal plasmids, but
it's fairly easy to sort through internals and externals (with plant DNA) with
simple mini-preps.

                    June Medford
                    jim2 at psuvm

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