ASSEMBLY OF HIGH-RESOLUTION BACTERIAL ARTIFICIAL CHROMOSOME, P1-DERIVED ARTIFICIAL CHROMOSOME, AND COSMID CONTIGS

The generation of contiguous physical maps is often complicated by a v ariety of factors including the type of cloning system used, Here we d escribe procedures for the isolation, rapid characterization, and phys ical mapping of large-insert recombinant bacterial clones from total h uman genomic BAC (bacterial artificial chromosome) and PAC (P1-derived artificial chromosome) libraries containing clones with an average in sert size of 150 kbp, After initial isolation, the clones were subject ed to a variety of fingerprinting procedures including inter-Alu PCR, semiautomated fluorescent fingerprinting, and EcoRI restriction fragme nt mapping, Individual BAC and PAC clones were also used as probes to interrogate arrayed chromosome 19-specific cosmid libraries, The combi nation of analyses facilitated the identification of chromosome-specif ic large-insert clones as well as the construction of a large (1.2 Mb) high-resolution BAG, PAC, and cosmid contig in 19q13.2, spanning the region from the carcinoembryonic antigen gene family to the X-ray repa ir cross complementing 1 DNA repair gene, This type of approach direct ly demonstrates the utility of large-insert recombinant bacterial clon es for the construction of contiguous physical maps of entire chromoso mes, (C) 1995 Academic Press,Inc.