| CHORI-82
Canine BAC Library |
 |
The “CHORI-82”
Canine BAC library has been constructed by Baoli Zhu in Pieter
de Jong’s laboratory at BACPAC Resources, Children’s
Hospital Oakland Research Institute. The preparation of the
library followed the cloning approach developed in our laboratory
(Osoegawa
et al., 1998). DNA was isolated from white blood cells
obtained from a female dog named Tasha (breed: Boxer; family
name: Canis familiaris). The sample was kindly provided by
Gregory M. Acland in the Center for Canine Genetics &
Reproduction, James A. Baker Institute for Animal Health,
NYS College of Veterinary Medicine Cornell University, Hungerford
Hill Rd, Ithaca, New York 14853 6401. High-molecular-weight
DNA was isolated from white blood cells and was embedded in
agarose to stabilize the DNA. The embedded DNA was partially
digested with a combination of EcoRI restriction enzyme and
EcoRI methylase and size fractionated by pulsed-field electrophoresis.
DNA fragments from the appropriate size fraction were cloned
into the pTARBAC2.1 vector between the two EcoRI sites. The
ligation products were transformed into DH10B (T1 resistant)
electro-competent cells (Invitrogen). The library will be
arrayed into 528 384-well microtiter dishes and will subsequently
be gridded onto 11 22x22cm
nylon high-density filters for screening by probe hybridization.
Each hybridization membrane represents over 18,000 distinct
BAC clones, stamped in duplicate. BAC library construction
was funded as part of the NIH-funded BAC Resource Network
by a grant from NIH (#HG025323-02).
Provisional Data for CHORI-82 Canine BAC library:
| Segment |
1 |
2 |
Total |
| Cloning Vector |
pTARBAC2.1 |
pTARBAC2.1 |
|
| DNA Source |
White blood cells |
White blood cells |
|
| Restriction Enzyme |
EcoRI |
EcoRI |
|
| Plate Range |
1-288 |
289-528 |
1-528 |
| Total 384-Well Dishes |
288 |
240 |
528 |
| Empty Wells(%) |
N/A |
N/A |
N/A |
| Non-insert Clones (%) |
N/A |
N/A |
N/A |
| Vector Clones (%) |
N/A |
N/A |
N/A |
| Recombinant Clones |
N/A |
N/A |
N/A |
| Insert size (Average) |
~185 Kbp |
~185 Kbp |
~185Kbp |
| Standard Deviation |
N/A |
N/A |
N/A |
| Library Redundancy (3.3 Gbp genome) |
~5.5 fold |
~4.5 fold |
~10 fold |
The CHORI-82 clone average insert size has been determined
from a sample of 172 colonies prior to the arraying of the
BAC library. Pulsed Field Gel Electrophoresis of Not1 digested
BACs with appropriate size standard was used for size determination.
Clone size distribution will
be posted on this page as soon
as available. 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).
N/A
|
