CONSTRUCTION AND CHARACTERIZATION OF HUMAN-CHROMOSOME 2-SPECIFIC COSMID, FOSMID, AND PAC CLONE LIBRARIES

Three human chromosome a-specific clone libraries were constructed and characterized. Chromosome 2-specific cosmid and fosmid clone librarie s were constructed using flow-sorted DNA from the monochromosomal hybr id cell line GM10826. The cosmid and fosmid libraries consist of 38,49 6 and 26,400 arrayed clones, respectively, with an average size of 40 kb. Colony hybridization of a representative number of clones with bot h human and hamster genomic DNA probes demonstrates that between 58 an d 66% of the clones in the how-sorted libraries contain human inserts. Approximately 5% of the cosmid and fosmid clones are nonrecombinants. A chromosome a-specific PAC library was also produced from the hybrid cell line GM10826. DNA from the hybrid cell line was cloned, and the human chromosome a-specific clones were identified by colony hybridiza tion. Approximately 5800 chromosome 2-specific PAC clones with an aver age insert size of similar to 85 kb were arrayed. Based on the size of the clones, the cosmid, fosmid, and PAC libraries are similar to 3.6x , similar to 2.5x, and similar to 1.9x, respectively, in chromosomal c overage. The chromosome 2 coverage of each of the three libraries was further determined by PCR screening clone pools with 82 chromosome 2-s pecific STSs. The average number of clones identified for each STS in the library indicates the cosmid, fosmid, and PAC libraries to be simi lar to 3.2x, similar to 2.1x, and similar to 1.5x, respectively, in ch romosome coverage. All except one of the 82 STSs were represented in t he portions of the libraries screened. (C) 1996 Academic Press, Inc.