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Problems with Biotin!

Chris Earl cearl at ra.isisnet.com
Mon May 20 15:13:38 EST 1996


Hello,

        I am a research technician, working on the amphibian nervous
system, and how it changes during metamorphosis.

        I am using the "BDA-10,000 Neuronal tracer kit (N-7167)" from
Molecular Probes Inc..  This kit contains biotin dextran amine,
avidin-horseradish peroxidase and DAB.

        What I have tried to do is take the vagus nerve after it exits
the jugular foramen, and with forceps crush the nreve and then apply
the Biotin.  However, this did not work.  NOw I know that the biotin
will travel anterogradely - but when I completed the reaction with the
avidin-HRP and DAB, there were not color changes or the typical brown
marks.  I did keep the application site and there was also nothing
there.

        Would anyone have suggestions on what I might be doing wrong
or another way I should go by applying the Biotin?

        I have thought of using microinjections - but I am trying to
keep this as a last resort - seeing as the vagus in the tadpole is
very small.

        Any suggestions and comments are welcome.

        Thank you in advance.

        Nathalie Major

please send replies to: nmajor at is.dal.ca





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