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quick mapping

Fri Apr 3 15:34:35 EST 1992

I am developing a quick mapping procedure which should facilitate
the process of mapping a gene to one of the ten Arabidopsis
chromosome arms.

In brief the method follows the segregation patterns of strain
specific PCR markers generated by primers based on the sequence
of previously mapped genes. Ideally, a collection of of primers
should correspond to sequences evenly distributed along the
chromosome arms.

I am in the process of identifying ecotype specific markers. One
class of such markers corresponds to ecotype specific insertions
or deletions.  At least 3 such sequences have been identified
serendipitously between Landsberg and Columbia ecotypes and I
hope to find more by genomic subtraction.  A second class of
markers will be generated by a combination of PCR amplification
of a previously sequenced region followed by restriction enzyme
digestion to display an RFLP.  

I would greatly appreciate information from anyone who has both
mapped and sequenced an Arabidopsis gene. For each mapped locus,
I will need about 1 kb of sequence.  In particular, I am short of
sequences mapped to chromosome 2 (there is only one available in
the Genbank).  Also if anyone encounters an ecotype-specific
deletion or insertion (e.g. during chromosome walking), I would
appreciate it if you make it available to me.  I will sequence it
(at least some of it) for you and use the sequence to generate
additional PCR primers.

As soon as I have identified a suitable set of mapping primers
and have identified restriction enzymes that reveal polymorphisms
in the PCR products, I will make this information avialable on
the network.

Thanks very much for your cooperation.

Andrzej Konieczny
Department of Molecular Biology
Massachusetts General Hospital.

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