|
CHORI-240: Bovine (Male) (Bull Li Domino) BAC Library |
 |
The CHORI-240 BAC
library has been constructed by Chung Li Shu and Kazutoyo
Osoegawa in our laboratory. Genomic DNA was isolated from
Hereford bull L1 Domino 99375 and was partially digested with
MboI. Size selected DNA was cloned into the pTARBAC1.3 vector
between the BamHI sites. The ligation products were transformed
into DH10B electrocompetent cells (BRL Life Technologies)
for segment 1 and DH10B T1 phage resistant electrocompetent
cells (BRL Life Technologies) for segment 2, respectively.
The library has been arrayed into 384-well microtiter dishes
and also gridded onto 11 22x22cm nylon
high-density filters for screening by probe hybridization.
Each hybridization membrane represents over 18,000 distinct
bovine BAC clones, stamped in duplicate. BAC library construction
was funded by a grant from USDA (99-35205-8537).
Blood was collected
from Hereford bull L1 Domino 99375 in Dr. Michael MacNeil's
laboratory, USDA-ARS, Miles City, MT. For additional information
with respect to the bovine strain, please contact Dr. Michael
MacNeil by email ([email protected]).
| Segment |
|
DNA |
Restriction Enzyme |
Plate Numbers |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC1.3 |
Blood |
MboI |
1-288 |
288 |
2,160 |
2.0 |
| 2 |
pTARBAC1.3 |
Blood |
MboI |
289-528 |
240 |
1,207 |
1.3 |
| Total |
|
|
|
1-528 |
528 |
3,367 |
1.7 |
| Segment |
Non-insert Clones
(%) |
Non-insert clones |
|
Recombinant Clones |
Insert size (Average) |
Redundancy (Genome
size:
3000 Mb) |
| 1 |
5.7 |
Approx. 6,180 |
0 |
Approx. 102,252 |
180 Kbp |
6.1 |
| 2 |
1.6 |
Approx. 1,455 |
15 |
Approx. 89,483 |
153 Kbp |
4.6 |
| Total |
|
|
15 |
Approx. 191,735 |
167 Kbp |
10.7 |
Data on the CHORI-240 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 .
|
