The RPCI-86 Male Feline BAC Library has been constructed at RPCI by Yu Wang. The project was funded by Ralston Purina from a grant proposal by Leslie A. Lyons, Ph.D., at The Laboratory of Genomic Diversity in the National Cancer Institute. The library was made from a random bred male domestic short-haired cat, a brown classic tabby. Blood was collected 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 22x22 cm high-density nylon hybridization filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct feline BAC clones, stamped in duplicate
The Total RPCI-86 Feline Library:
| Segment | Cloning Vector | DNA | Plate Numbers | Total Plates | Total Clones | Empty Wells (Total) |
| 1 | pTARBAC2 | Cat White Blood Cell | 1-288 | 288 | 107,041 | 781 |
| 2 | pTARBAC2 | Cat White Blood Cell | 289-624 | 336 | 127,308 | 1,320 |
| Total Library | 1-624 | 624 | 234,349 | 2,101 |
| Segment | Empty Wells (%) | Non-Recombinant Clones (Total) | Non-Recombinant Clones (%) | Insert Size (average) | Genomic Coverage |
| 1 | 0.7 | approx. 2,770 | 2.5 | 138 Kbp | 4.8x |
| 2 | 1.0 | approx. 780 | 0.6 | 137 Kbp | 5.8x |
| Total Library | 0.9 | approx. 3,550 | 1.5 | 137 Kbp | 10.6x |
click here for a legend of the previous tables.
The average insert size data has been determined by Pulsed Field Gel Electrophoretic analysis of clones. Further (in depth) characterization of the library is ongoing in our lab and data will be updated on our WebPage periodically.