[Zbrafish] Re: Tol2 injections

Wilson Clements via zbrafish%40net.bio.net (by wclements from ucsd.edu)
Tue Jul 22 19:55:16 EST 2008


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
------------------------------------------------------------------------ 
--------------------
Wilson Clements, Ph.D.

wclements from ucsd.edu

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


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