SERVICE UPDATE AT ZIRC
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Sources for Probes Not Distributed by ZIRC
Two BAC libraries are currently available that have been
Tuebingen strain DNA, the same DNA currently
be used for the
Sanger Center genome
CHORI-211 BAC library
The CHORI-211 BAC library was constructed by Chung
Li Shu and Kazutoyo Osoegawa in Pieter J. de Jong’s
laboratory at Children's Hospital Oakland Research
Institute. Genomic DNA was isolated from testis
of 6 males by Dr. Gerd-Joerg Rauch and partially
digested with a combination of EcoRI and EcoRI Methylase.
Size selected DNA was cloned into the pTARBAC2.1
vector between the EcoRI sites. The ligation products
were transformed into DH10B electro-competent cells
(BRL Life Technologies). The library has been arrayed
into 384-well microtiter dishes and also gridded
onto 6 22x22cm nylon high-density filters for screening
by probe hybridization. Each hybridization membrane
represents over 18,000 distinct zebrafish BAC clones,
stamped in duplicate. Library construction was supported
by sub-contracts from a grant awarded to
at The Max-Planck-Institut für Entwicklungsbiologie,
Tübingen. For more information check:
Daniokey BAC library
The Daniokey BAC library, created by the
Laboratory and Keygene N.V., is available
via the RZPD
in Berlin. The library is available as glycerol stocks,
filters, and PCR pools. It, too, was constructed
genomic DNA. A partial HindIII digest
resulted in 104,064 BAC clones with an average insert
size of ~175 kb. The library provides approximately
10x coverage of the zebrafish genome.
Other BAC libraries used in the Sanger project.
BUSM1 PAC library
BUSM1 PAC library
was constructed by
while he was at Boston University School of Medicine. The library
was constructed using MboI partial digests of high molecular weight DNA
from blood cells of ~ 200 individuals (
) ligated to the BamHI sites of pCYPAC6 (GenBank AF133437). The
library is archived in 271 384-well dishes. High-density filters and
PCR-screenable pools for this library are available from the
In addition to the RZPD, numerous other labs have replicas of this library,
Will Talbot, Steve Johnson, John Postlethwait, Len Zon, Marnie Halpern,
Shuo Lin, and Didier Stanier. Copies of the library also exist in Japan
and Europe. Special requests for pools for PCR screening and/or high
density filters can be made to Chris Amemiya (email@example.com).
Zebrafish YAC library
Zebrafish YAC library,
constructed by the Massachusetts General Hospital in collaboration
with the MIT Center for Genome Research, is available from
The average insert size for the library is 480 kb. With approximately
19,000 clones, the library represents five zebrafish genome equivalents.
Inserts were cloned into the EcoRI site of the pRML1 and pRML2 vector arms.
J57D was used as the yeast host strain. Ref: T.P. Zhong, K. Kaphingst,
U.Akella, M. Haldi, E.S. Lander and M. Fishman Genomics 48, 136-138 (1998).
Random-primed shield-stage cDNA library
A random-primed shield-stage cDNA library, cloned into lambda ZAPII, and
amplified from 6.5x106 independent recombinants is available
Laboratory of Molecular Genetics
Building 6B, Room 4B-422
9000 Rockville Pike
Bethesda, MD 20892
fax: (301) 496-0243
Embryonic cDNA libraries are unfortunately no longer available from
David Grunwald or Bruce Appel.
If you know of any other sources, please