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CHORI-242: Porcine (Sus scrofa) BAC Library |
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The CHORI-242 porcine
(Sus scrofa) 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). Blood sample was obtained from a single
Duroc female (Herd ID 2-14) located at the UIUC Moorman Swine
Research farm through Dr. Jonathan E. Beever (Department of
Animal Sciences, University of Illinois at Urbana-Champaign).
High-molecular-weight DNA was isolated from white blood cells.
Agarose embedded high molecular weight DNA was partially digested
with MboI and size fractionated DNA was cloned into the pTARBAC1.3
vector between the BamHI sites. The ligation products were
transformed into DH10B (T1 resistant) electrocompetent cells
(Invitrogen). The library has been arrayed into 384-well microtiter
dishes and also gridded onto eleven 22x22cm
nylon high-density filters for screening by probe hybridization.
Each hybridization membrane represents over 18,000 distinct
porcine BAC clones, stamped in duplicate. Library characterization
was performed by Qing Cao and Kazutoyo Osoegawa. BAC library
construction was funded by a grant from USDA (99-35205-8537).
Provisional
data for CHORI-242 Procine BAC Library:
| Segment |
Cloning Vector |
DNA |
Restriction enzyme |
Plate Number |
Total Plates |
Empty Wells |
Empty Wells (%) |
| 1 |
pTARBAC1.3 |
White Blood Cell |
MboI |
1-288 |
288 |
1,126 |
1.02 |
| 2 |
pTARBAC1.3 |
White Blood Cells |
MboI |
289-528 |
240 |
952 |
1.03 |
| Total Library |
|
|
|
1-528 |
528 |
2,078 |
1.02 |
| Segment |
Non-insert clones (%) |
Non-insert clones |
Non-Recombinant Clones (pUC) |
Recombinant clones |
Insert Size (Average) |
Redundancy (Genome Size: 3000 Mb) |
| 1 |
2.08 |
Approx. 2,300 |
0 |
Approx. 107,166 |
177Kbp |
6.3 |
| 2 |
2.08 |
Apporx. 1,917 |
0 |
Approx. 89,291 |
170Kbp |
5.1 |
| Total Library |
2.08 |
Apporx. 4,217 |
0 |
Approx. 196,457 |
173Kbp |
11.4 |
Data on the CHORI-242 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.
Please direct questions
concerning this library to either Pieter
J. de Jong .
| 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 ( [email protected]
), fax: (510) 450-7924).
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