Zebrafish BAC clones
For zebrafish researchers, BACs may also be useful starting points for transgene construction. Although germline transgenesis of full-length BACs is less efficient in fish than in mice, recent developments such as Tol2 transposon-mediated transgenesis may help (Suster et al. 2009).
1. Workflow for using fish BACs
A typical workflow of recombineering and injecting BACs might be as follows:
a. Transfer BAC into strain permitting homologous recombination.
b. Recombineer the BAC as desired.
c. Purify BAC DNA by cesium choride gradient or BAC column kit.
However, the cesium chloride method generally does not yield BAC DNA in sufficient quantity for injection in fish embryos.
d. Verify concentration and integrity of purified BAC DNA by agarose gel analysis.
e. Dilute BAC DNA prior to injection.
2. Where to get zebrafish BACs
http://bacpac.chori.org/ . The BACPAC Resources Center is located at the
Children's Hospital Oakland Research Institute (CHORI), Oakland, CA.
RPCI-71: Zebrafish ( Danio rerio ) BAC Library
CHORI-73: Doubled-haploid Zebrafish (Tuebingen line) ( Danio rerio ) BAC Library
CHORI-211: Zebrafish Tue (Tuebingen) strain ( Danio rerio ) BAC Library
CHORI-1073: Doubled haploid Zebrafish ( Danio rerio, Tuebingen line ) Fosmid Library
DKEY (DanioKey) BAC library:
c.
age : “
How can I find a BAC/PAC clone?” http://www.sanger.ac.uk/Projects/D_rerio/faqs.shtml
Doug Mortlock 2011/2012