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Successful backcrosses

Meisel, Lee A meisel at OCELOT.RUTGERS.EDU
Wed Sep 23 18:51:47 EST 1992

         Hello!   I am a graduate student at Rutgers university studying the
molecular basis of leaf morphology in Arabidopsis.  Recently many papers,
review articles and even books have been written about Arabidopsis and the many
advantages associated with this small plant.
Testerlines, RFLP maps and many known mutants are available for us to
characterize new mutants as well as chromosomally locate these mutants.  
However,  to my dismay, I have not been able to locate information on the
technical aspects of performing backcrosses and crosses with the various tester
and mutant lines available.
         If anyone out there in Arabidopsis land could supply me with technical
 information (published or unpublished) on crosses it would be very beneficial
to me as well as my project.
         Presently, I am performing the crosses under a  mechanical drawing
lamp (a magnifying glass surrounded by a flourescent light).  This allows me to
emassculate the floral buds without harming the carpel and my eyes.  After 
emassculation, I cover the inflorescence with a small grain bag supported by a
plastic coffee stirrer.  After approximately two days I pollinate the
emassculated carpel swiping the anthers of the male donor over the carpel. 
This has enabled me to successfully isolate  unpollenated carpels, but I am
having problems at the pollination step.

            Does any one know when the pollen grains are released from the
anthers and at what stage of floral development the pollen grains are most
viable (is it before the flower opens as it is in Mendel's peas) ?

              Could any of you who perform crosses on a regular basis provide
me with tips on how to optimize my crosses?

              Is the release of pollen grains in Arabidopsis dependent upon
light or temperature (another words is it necessary to pollenate the
emassculated carpels early in the morning to increase your success rate) ?

    I realize crosses in Arabidopsis are done routinely, however being new to
the Arabidopsis field I am unfamiliar with the basic technical aspects of such
crosses.  I would appreciate any information you could provide me.

Thank you 
                    Lee Meisel
                    Waksman Institute
                    Piscataway, NJ  08855-0759

                  Phone # (908)932-4296
                  Fax # (908)932-5735
                  email =  meisel at mbcl.rutgers.edu 

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