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 Home > Resources > Libraries
   RPCI - 23 Female (C57BL/6J) Mouse BAC Library   

The RPCI-23 BAC Library has been constructed in our laboratory by Kazutoyo Osoegawa and Minako Tateno* from pooled tissues derived from three 5-week old female C57BL/6J mice (obtained from the Jackson Laboratory). Mouse kidney and brain genomic DNA samples were isolated and partially digested with a combination of EcoRI and EcoRI Methylase. Size-selected EcoRI fragments were cloned in 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-23 C57BL/6J Mouse BAC Library:

Segments Cloning Vector DNA Plate Numbers Total Plates Total Clones Empty Wells (Total)
1 pBACe3.6 Female (kidney/brain) 1-240 240 90,443 1717
2 pBACe3.6 Female (kidney/brain) 241-480 240 89,953 2207
Total Library       480 180,396 3924

 

Segments % Empty Wells 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. 

The library is available for individual clone orders, distributed as agar-stab cultures at a not-for profit cost. See our webpage "Ordering & Pricing Information" for the costs.

The BAC library has been gridded onto 22x22cm positively charged nylon filters for hybridization screening purposes. Each filter contains 36,864 colonies which represents 18,432 independent clones spotted in duplicate in a 4x4 clone array. Clones can be identified through screening by purchasing "high-density colony" hybridization filters or by utilizing our "fee for service" "Library Screening Services" . For questions about clones and hybridization membranes, please contact BACPAC Resources (BACPACorders@chori.org).

*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

With respect to the scheduling of shipments of arrayed library copies, we would like to make you aware of our policy for array distribution.

Please visit the ordering information page if you want to place an order.

Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong (pdejong@chori.org, fax: (510) 450-7924). For hybridization membranes, please contact BACPAC Resources (BACPACorders@chori.org).

 

 

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