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 variety of factors including the type of cloning system used. Here we describe procedures for the isolation, rapid characterization, and physical mapping of large-insert recombinant bacterial clones from total human genomic BAC (bacterial artificial chromosome) and PAC (P1-derived artificial chromosome) libraries containing clones with an average insert size of 150 kbp. After initial isolation, the clones were subjected to a variety of fingerprinting procedures including inter-Alu PCR, semiautomated fluorescent fingerprinting, and EcoRI restriction fragment mapping. Individual BAC and PAC clones were also used as probes to interrogate arrayed chromosome 19-specific cosmid libraries. The combination of analyses facilitated the identification of chromosome-specific large-insert clones as well as the construction of a large (1.2 Mb) high-resolution BAC, PAC, and cosmid contig in 19q13.2, spanning the region from the carcinoembryonic antigen gene family to the X-ray repair cross complementing 1 DNA repair gene. This type of approach directly demonstrates the utility of large-insert recombinant bacterial clones for the construction of contiguous physical maps of entire chromosomes. © 1995 Academic Press, Inc.
Publication Source (Journal or Book title)
Ashworth, L., Alegria-Hartman, M., Burgin, M., Devlin, L., Carrano, A., & Batzer, M. (1995). Assembly of high-resolution bacterial artificial chromosome, P1-derived artificial chromosome, and cosmid contigs. Analytical Biochemistry, 224 (2), 564-571. https://doi.org/10.1006/abio.1995.1088