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 Home > Resources > Libraries
  RPCI-24 Mouse (C57BL/6J Male) BAC Library   

The RPCI-24 BAC library has been constructed in our laboratory from tissues derived from single male C57BL/6J mouse (obtained from the Jackson Laboratory). Spleen and brain genomic DNA samples were isolated and partially digested with MboI. Size-selected MboI fragments were cloned into the pTARBAC1 vector between the BamHI 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-24 Male (C57BL/6J) Mouse BAC Library

Segments Cloning Vector DNA Restriction
enzyme
Plate Numbers Total Plates Empty Wells Empty Wells (%)
1 pTARBAC1 Spleen MboI 1-161, 168-177
288 3464 3.1
Brain MboI 162-167, 178-288
2 pTARBAC1 Brain MboI 289-313 288 3951 3.6
Spleen MboI 314-576
Total Library         576 7415 3.35

 

Segments Non-insert Clones (%) Non-insert clones Non-Recombinant Clones (pUC) Recombinant Clones Average Insert Size
Redundancy
(Genome size:
3,000 Mb)
1 4.3 4607 1 102,521 160 Kbp 5.5
2 0.5 533 0 106,108 150 Kbp 5.3
Total Library 2.4 5140 1 208,629 155 Kbp 10.8

click here for a legend of the previous tables.


Data on the RPCI-24 clone average insert size (Segment 1, Segment 2) has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically.

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).

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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