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CHORI-241: Equine (Equus caballus) BAC Library |
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The CHORI-241 Horse
(Equus caballus) BAC library has been generated
by Baoli Zhu, in Pieter de Jong's laboratory at BACPAC Resources,
Children's Hospital Oakland Research Institute. The library
preparation followed the general cloning approach developed
in our laboratory Osoegawa
et al., 1998 . DNA was obtained from a blood sample provided
by Dr. Doug Antczak of the Equine Genetic Center, James A.
Baker Institute for Animal Health, College of Veterinary Medicine,
Cornell University. The donor of this sample was a thoroughbred
male horse by the name "Bravo", a half-brother of
the female "Twilight" used for horse genome sequencing
as part of the NIH horse
genome project. A news release related to the project
and the horses involved in the "BAC library" and
"Horse Genome" Projects can be found in the online
archive of The Cornell Daily Sun (http://cornellsun.com/node/21536).
High molecular weight DNA was prepared by embedding peripheral
blood neutrophil cells in an agarose matrix. (done by Dr.
Helen Bird at Cornell). The library has been constructed by
using partially digested DNA with EcoRI plus EcoRI methylase.
Size fractionated DNA was cloned into the pTARBAC2.1 vector
between the EcoRI sites. The ligation products were transformed
into DH10B eletrocompetent cells (BRL Life Technologies).
The library has been arrayed into 384-well microtiter dishes
and also gridded onto a 22x22cm nylon high-density filter
for screening by probe hybridization.
| Segments |
Cloning Vector |
DNA |
Restriction
enzyme |
Plate Numbers |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC2.1 |
Male Neutrophils |
EcoRI |
1-192 |
192 |
1,622 |
2.2% |
| 2 |
pTARBAC2.1 |
Male Neutrophils |
EcoRI |
193-528 |
336 |
2,838 |
2.2% |
| Total Library |
|
|
|
1-528 |
528 |
4,460 |
2.2% |
| Segments |
Non-insert Clones (%) |
Non-insert clones |
Non-Recombinant Clones (pUC) |
Recombinant Clones |
Average Insert Size |
Redundancy
(Genome size:
3.0 Mb)
|
| 1 |
2.1% |
Approx. 1,514 |
552 |
Approx. 70,040 |
171Kbp |
4.0 |
| 2 |
3.9% |
Approx. 4,921
|
653 |
Approx. 120,612 |
171Kbp |
6.9 |
| Total Library |
3.3% |
Approx. 6,435 |
1,205 |
Approx. 190,652
|
171Kbp |
10.9 |
The total library
should represent approximately 11.8-fold total genomic representation.
Data on the CHORI-241
clone average insert size has been determined by Pulsed
Field Gel Electrophoresis. Clone size distribution 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.
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 ( [email protected]
), fax: (510) 450-7924).
Please direct questions
concerning this library to either Pieter
J. de Jong .
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