RPCI - 23 Female (C57BL/6J) Mouse BAC Library
The RPCI-23 BAC Library has been constructed in our laboratory by Kazutoyo Osegawa and Minako Tateno*. Female C57BL/6J mouse kidney and/or brain genomic DNA was isolated and partially digested with a combination of EcoRI and EcoRI Methylase. Size selected DNA was cloned between the pBACe3.6 vector at 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 mouse BAC clones, stamped in duplicate.
The RPCI-23 C57BL/6J Mouse BAC Library:
| Segments | Cloning Vector | DNA | Plate Numbers | Total Plates | Total Clones | Empty Walls (Total) |
| 1 | pBACe3.6 | Female (kidney/brain) | 1-240 | 240 | 80661 | 1717 |
| 2 | pBACe3.6 | Female (kidney/brain) | 241-480 | 240 | 89973 | 2207 |
| Total Library | 480 | 170634 | 3924 |
| Segments | % Empty Walls | Non-Recombinant Clones (Total) | Non-Recombinant Clones (%) | Insert Size (average) | Genomic Coverage |
| 1 | 1.9 | approx. 10523 | 11.3 | 198 Kbp | 5.3X |
| 2 | 2.4 | approx. 1211 | 1.3 | 197 Kbp | 5.9X |
| Total Library | 2.1 | approx. 11734 | 6.8 | 197 Kbp | 11.2X |
click here for a legend of the previous tables.
Data on the RPCI-23 clone average insert size
has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution
has been plotted graphically.
Reference:
Osoegawa, K. et al. (2000) Bacterial Artificial Chromosome Libraries for Mouse
Sequencing and Functional Analysis. Genome Research 10: 116-128
Reprints availiable upon request.
*Visiting scientist supported by the RIKEN Institute, permanent address:
Minako Tateno
RIKEN Tsukuba Life Science Center
Genome Science Laboratory
3-1-1 Koyadai Tsukuba, Ibaraki 305
Japan