RPCI - 22 (129S6/SvEvTac) Mouse BAC Library
The RPCI-22 BAC Library has been constructed in our laboratory by Kazutoyo Osoegawa. Female 129S6/SvEvTac (Taconic) mouse spleen genomic 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 mouse BAC clones, stamped in duplicate.
The RPCI - 22 129S6/SvEvTac Mouse BAC Library:
Segment |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
|
| 1 | pBACe3.6 | Female Spleen | 1-288 | 288 | 102,914 | 1,678 |
| 2 | pBACe3.6 | Female Spleen | 289-576 | 288 | 109,039 | 1,443 |
| Total Library | 576 | 211,953 | 3,121 |
| Segment | Empty Wells (%) | Non-Recombinant Clones (Total) | Non-Recombinant Clones (%) | Average Insert Size | Genomic Coverage |
| 1 | 1.5 | approx. 6,000 | 5.5 | 155 Kbp | 5.3X |
| 2 | 1.3 | approx. 110 | 0.1 | 154 Kbp | 5.6X |
| Total Library | 1.4 | approx. 6,110 | 2.9 | 154 Kbp | 10.9X |
click here for a legend of the previous tables.
Data on the RPCI-22 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.