| CHORI-241
Equine BAC Library
|
 |
The CHORI-241 Horse
(Equus caballus) BAC library has been constructed
by Dr. Baoli Zhu in BACPAC Resources in collaboration with
Dr. Doug Antczak in Equine Genetic Center, James A. Baker
Institute for Animal Health, College of Veterinary Medicine,
Cornell University. High molecular weight DNA was prepared
in embedded agarose plugs from thoroughbred male peripheral
blood neutrophil cells by Dr. Helen Bird. 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 ( pdejong@chori.org
), fax: (510) 450-7924).
Please direct questions
concerning this library to either Pieter
J. de Jong or Kazutoyo
Osoegawa .
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