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 Home > Resources > Libraries
  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

Cloning Vector

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.

The library is available as library plates, individual clones, and high-density hybridization sets (see Filter Availability)

Individual clones identified after screening can be obtained as stab cultures at a cost of $80 per clone in addition to a $35 set-up charge per shipment. Shipping will be done by Federal Express and charged to the recipients account number.

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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