RPCI Human PAC Library

RPCI Human PAC libraries have been constructed in our laboratory by Panos A. Ioannou* (RPCI-1), Chira Chen and Baohui Zhao (RPCI-3), and Baohui Zhao (RPCI-4, 5, and 6).   Library construction was funded by grants form the U.S. Department of Energy (#DE-FG02-94ER61883) and the National Human Genome Research Institute (NHGRI, NIH) (#1R01RG01165).  Anonymous male (RPCI-1, 3, 4, AND 5) or female (RPCI-6) blood DNA was isolated, partially digested with MboI and cloned between the BamH1 sites of the pCYPAC2 (RPCI-1, 2, 3, 4, and 5) or pPAC4 (RPCI-6) vector.  The ligation products were transformed into DH10B electrocompetent cells (BRL Life technologies).  The libraries have been arrayed into 384-well microtiter dishes and gridded onto 22x22cm nylon high density hybridization filters for screening purposes.

*Present Address:
        Panos Ioannu, Ph.D.
        The Murdoch Institute for Research into Birth Defects
        Royal Children's Hospital
        Flemington Road, Parkville 3052
        Melbourne, Australia

The Library segments have been characterized as to:

    - The percentage of empty wells
    - Percentage of non-recombinant clones
    - Average insert size
    - Approx. genomic coverage

The clone size distribution for each segment of the library has also been determined.