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 Home > Resources > Libraries
  RPCI - 86 Male Feline BAC

  

The RPCI-86 Male Feline BAC Library has been constructed in our laboratory by Yu Wang.  The project was funded by Ralston Purina from a grant proposal by Leslie A. Lyons, Ph.D., at The Laboratory of Genomic Diversity in the National Cancer Institute.  The library was made from a random bred male domestic short-haired cat, a brown classic tabby.  Blood was collected and genomic DNA was isolated from the white blood cells.  The DNA (partially EcoRI digested) was size selected and cloned between the EcoRI sites of the pTARBAC2  vector.  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 22x22 cm high-density nylon hybridization filters for screening by probe hybridization.  Each hybridization membrane represents over 18,000 distinct feline BAC clones, stamped in duplicate

 

The Total RPCI-86 Feline Library:

Segment Cloning Vector DNA Plate Numbers Total Plates Total Clones Empty Wells (Total)
1 pTARBAC2 Cat White Blood Cell 1-288 288 107,041 781
2 pTARBAC2 Cat White Blood Cell 289-624 336 127,308 1,320
Total Library     1-624 624 234,349 2,101

         

Segment Empty Wells (%) Non-Recombinant Clones (Total) Non-Recombinant Clones (%) Insert Size (average) Genomic Coverage
1 0.7 approx. 2,770 2.5 138 Kbp 4.8x
2 1.0 approx. 780 0.6 137 Kbp 5.8x
Total Library 0.9 approx. 3,550 1.5 137 Kbp 10.6x

click here for a legend of the previous tables.

The average insert size data has been determined by Pulsed Field Gel Electrophoretic analysis of clones.  Further (in depth) characterization of the library is ongoing in our lab and data will be updated on our WebPage periodically.

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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