| RPCI
- 44 Male Porcine BAC Library
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The RPCI-44 Male Porcine BAC Library has been constructed
in our laboratory by Chung-Li Shu. Library construction
was supported by a contract from the United States Department
of Agriculture, Agricultural Research Service, U.S. Meat Animal
Research Center, Clay Center, NE. Blood from four male
pigs (breed: 37.5% Yorkshire, 37.5% Landrace, and 25% Meishan)
was pooled and genomic DNA was isolated from the white blood
cells. The DNA (partially EcoRI digested) was size selected
and cloned between the EcoRI sites of the pTARBAC2 vector.
The ligation products were transformed into DH10B electrocompetent
cells (BRL Life Technologies). The library has been
arrayed into 384-well microtiter dishes and also gridded onto
22x22cm
high-density nylon hybridization filters for screening
by probe hybridization. Each hybridization membrane
represents over 18,000 distinct porcine BAC clones, stamped
in duplicate.
The Total RPCI-44 Male Porcine BAC Library:
| Segment |
Cloning
Vector |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Total Empty Wells |
| 1 |
pTARBAC2 |
Pig* White Blood Cell |
1-240 |
240 |
83,946 |
1,764 |
| 2 |
pTARBAC2 |
Pig* White Blood Cell |
241-528 |
288 |
101,443 |
1,519 |
| Total Library |
|
|
1-528 |
528 |
185,389 |
3,283 |
| Segment |
(%) Empty Wells |
Total Non-Recombinant
Clones |
% Non-Recombinant Clones |
Average Insert Size
(Kb) |
Genomic Coverage |
| 1 |
1.9 |
approx. 6,450 |
7.0 |
157 |
4.4X |
| 2 |
1.2 |
approx. 7,630 |
6.9 |
171 |
5.8X |
| Total Library |
1.6 |
approx. 14,080 |
7.0 |
165 |
10.2X |
click here
for a legend of the previous tables.
*The RPCI-44 BAC Library was constructed from pooled pig
(4 pigs total) blood. The pigs were males, Breed: 1/4
Meishan, 3/8 Yorkshire, and 3/8 Landrace.
The average
insert size data has been determined by Pulsed Field Gel
Electriphoretic analysis of clones randomly chosen from plates
from each segment. Further (in depth) characterization
of the library is ongoing in our lab and data will be updated
on our WebPages periodically.
The library is available as library plates, individual clones, and high-density
hybridization sets (see Filter Availability)
Individual clones identified after screening can be obtained as stab cultures
at a cost of $80 per clone in addition to a $35 set-up charge per shipment. Shipping will be done by Federal Express and charged to the recipients account
number.
With respect to the scheduling of shipments of arrayed library copies, we would
like to make you aware of our policy for
array distribution.
Please visit the ordering information
page if you want to place an order.
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). For hybridization
membranes, please contact BACPAC Resources (BACPACorders@chori.org).
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