The RPCI-44 Male Porcine BAC Library has been constructed at RPCI 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.