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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 ([email protected], fax: (510) 450-7924). For hybridization membranes, please contact BACPAC Resources ([email protected]).
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