Tol2 injections

Dear ZF Community,

We are trying to optimize our dosage for Tol2 transgenesis. I was
hoping to poll people who are using this system to see roughly what
amounts of transgenesis plasmid and Tol2 transposase mRNA you are
injecting to obtain transgenics.

For no particular reason, we started out using 50 pg of DNA and 50 pg
of RNA. We find we get high levels of transgenesis and many
insertions per genome. On the plus side, many insertions gives
higher expression in the F1. But on the negative side, it makes it
easier to see position effects, lose expression over successive
generations, and makes it difficult to track specific insertions. If
you have experiences you would be willing to share in optimizing your
injections to whatever parameters you feel are important please send
them either to the list server or to me directly at the e-mail
address below.

Best,
Wilson
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Wilson Clements, Ph.D.

[Only registered users see links. ]

Dept. of Biology
Section of Cell and Developmental Biology
University of California at San Diego
9500 Gilman Dr.
Natural Sciences Building 6105
La Jolla, CA 92093-0380

TEL (858) 534-6955
LAB (858) 822-4658
FAX (858) 822-5740