[Zbrafish] Zebrafish transgenics

[Burdine, Rebecca D]

Hi Tania,
 
I would suggest reading up on the TOL2 transposase method of making
transgenics.  Both Nathan Lawson and Chi-Bien Chen have great GATEWAY
vectors that utilize TOL2 that both labs are providing to the community.
 
With Tol2 you can minimize mosaicism in transient injections, but to
eliminate this entirely you need to make a stable line.
 
If you haven't already, I would go to www.zfin.org and see what stable
lines they have for sale that might suit your needs.  You can also look
up to above labs and go to their websites to request plasmids.
 
Becky
 
---------------------------------------------------
Rebecca D. Burdine, Ph.D.
Assistant Professor
Dept. of Molecular Biology
Princeton University
Washington Road Mof 433
Princeton, NJ 08544 
 
Phone: (609) 258-7515
Fax: (609) 258-1343
Email: rburdine from princeton.edu
Admin Assistant: Cathy Falk (609) 258-1604
 


________________________________

	From: zbrafish-bounces from oat.bio.indiana.edu
[mailto:zbrafish-bounces from oat.bio.indiana.edu] On Behalf Of Tania Vinagre
	Sent: Tuesday, September 25, 2007 2:19 PM
	To: zbrafish from magpie.bio.indiana.edu
	Subject: [Zbrafish] Zebrafish transgenics
	
	

	I work with mice and consequently I have some limitations
regarding numbers of transgenic embryos I can obtain. For this reason, I
am considering doing some experiments in zebrafish. I am interested in
getting a reasonably high number of transgenic embryos and analyzing
their skeletal phenotype. At the moment I am not interested in producing
stable lines, only embryos, but I came across a number of possible
problems. Is there any simple way to overcome the mosaicism in zebrafish
transgenics? How can I promote the integration of my construct? Is there
any good review on transgenic techniques? 

	Thanks in advance

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