The RPCI-42 Bovine Library has been constructed at RPCI By Chung-Li Shu with funding provided through the Monsanto Corporation. Holstein Bull white blood cell DNA was isolated and partially digested with a combination of EcoRI and EcoRI Methylase. Size selected DNA was cloned into the pBACe3.6 vector between the EcoRI sites. 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 nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct bovine BAC clones, stamped in duplicate.
The RPCI-42 Bovine BAC Library:
| Segment | Cloning Vector | DNA | Plate Numbers | Total Plates | Total Clones | Empty Wells (Total) |
| 1 | pBACe3.6 | Holstein Bull White Blood Cell | 1-288 | 288 | 108,776 | 1,156 |
| 2 | pBACe3.6 | Holstein Bull White Blood Cell | 289-576 | 288 | 107,663 | 1,379 |
| Total Library | 576 | 216,439 | 2,535 |
| Segment | Empty Wells (%) | Non-Recombinant Clones (Total) | Non-Recombinant Clones (%) | Average Insert Size | Genomic Coverage |
| 1 | 1.0 | approx. 660 | 0.6 | 165 Kbp | 6.0X |
| 2 | 1.2 | approx. 1,550 | 1.4 | 163 Kbp | 5.9X |
| Total Library | 1.1 | approx. 2,210 | 1.0 | 164 Kbp | 11.9X |
click here for a legend of the previous tables.
The total library should represent approximately 12-fold total genomic representation. The RPCI-42 Bovine BAC Library clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically. The RPCI-42 Bovine BAC Library is available both as library plates and high-density hybridization filter sets (see filter availability).