RPCI - 32 Male (Brown Norway) Rat BAC Library


The RPCI-32 Male Brown Norway Rat BAC Library has been constructed on our laboratory by Chung-Li Shu.  Library construction was supported by a sub-contract from the Whitehead Institute for Biomedical Research.  Funding was from a NIH grant (5R01HL55726-03) awarded to E. Lander (P.I.).  Male Brown Norway (BN/ss/NH/sd) rat spleen genomic DNA (partially EcoRI digested) was cloned into DH10B electrocompetent cells (BRL Life Technologies).  The library has been arrayed into 384-well microtiter dishes and also gridded onto 22x22cm high-density nylon hybridization filters for screening by probe hybridization.  Each hybridization.  Each hybridization membrane represents over 18,000 distinct rat BAC clones, stamped in duplicate.

 

The Total RPCI-32 Rat BAC Library:

Segment Cloning Vector DNA Plate Numbers Total Plates Total Clones Total Empty Wells
1 pBACe3.6 Male Brown Norway Rat Spleen 1-288 288 106,648 1,244
2 pBACe3.6 Male Brown Norway Rat Spleen 289-624 336 121,665 1,559
Total Library     1-624 624 228,313 2,803

 

Segment (%) Empty Wells Total Non-Recombinant Clones % Non-Recombinant Clones Average Insert  Size (Kb) Genomic Coverage
1 1.0 approx. 2,700 2.5 147 5.2X
2 1.2 approx. 5,800 4.5 153 6.2X
Total 1.1 approx. 8,500 3.7 150 11.4X

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 web page periodically.