| CHORI-211
Zebrafish BAC Library
|
 |
The
CHORI-211 BAC library has been constructed by Chung Li Shu
and Kazutoyo Osoegawa in our laboratory. 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 Dr. Robert Geisler at
The Max-Planck-Institut fuer Entwicklungsbiologie.
The
strain name of zebrafish is Tue (Tuebingen).
The characteristics of this fish line are described in the
following paper:
Rauch, G.-J., Granato,
M. and Haffter, P. (1997). A polymorphic zebrafish line for
genetic mapping using SSLPs on high-percentage agarose gels.
Technical Tips Online T01208.
Provisonal
data for CHORI-211 Zebrafish BAC Library:
| Segment |
|
DNA |
Restriction Enzyme |
Plate Numbers |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC2.1 |
Testis from 6 males |
EcoRI |
1-144 |
144 |
1,314 |
2.4 |
| 2 |
pTARBAC2.1 |
Testis from 6 males |
EcoRI |
144-288 |
144 |
1,278 |
2.3 |
| Segment |
Non-insert Clones
(%) |
Non-insert clones |
|
Recombinant clones |
Insert size (Average) |
Redundancy (Genome
size:
1,700 Mb) |
| 1 |
1.2 |
approx. 648 |
0 |
53,334 |
171 Kbp |
5.4 |
| 2 |
2.6 |
approx. 1404 |
41 |
52,573 |
160 Kbp |
5.0 |
Data on the CHORI-211
clone average insert size of Segment 1 and Segment
2 have been determined by Pulsed Field Gel Electrophoresis.
Clone size distribution for each segment has been plotted
graphically. While analyzing clones
using pulse-field electrophoresis to determine the average
insert size, non-insert clones containing a small deleted
vector fragment consistent with sucrose resistance were observed.
Further in depth characterization of the library is
on going in our lab and data will be updated on our web page
periodically.
Please direct questions
concerning this library to either Pieter
J. de Jong or Kazutoyo
Osoegawa.
| Ordering &
Pricing information |
The library is available
in several formats. Individual clones, and high-density hybridization
filters are obtainable. For ordering and shipping
details, please view the ordering
and pricing information page.
Academic and commercial
users interested in a copy of the BAC library should contact
Pieter J. de Jong ( pdejong@chori.org
), fax: (510) 450-7924).
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