Selection of chromosome 22-specific clones from human genomic BAC library using a chromosome-specific cosmid library pool

Genomics. 1994 Jul 15;22(2):336-9. doi: 10.1006/geno.1994.1392.

Abstract

A new approach to rapidly identify chromosome-specific subsets of clones from a total human genomic library is described. We report here the results of screening a human bacterial artificial chromosome (BAC) library using the total pool of clones from a chromosome 22-specific cosmid library as a composite probe. The human BAC library was gridded on filters at high density and hybridized with DNA from the pooled chromosome 22-specific Lawrist library under suppressive conditions. In a single hybridization, we picked 280 candidates from the BAC library representing over 30,000 clones (or 1.2x coverage of human genome). This subset contained more than 60% of the chromosome 22-specific BAC clones that were previously found to be present in the original BAC library. In principle, this approach can be applied to select a subset of clones from other global libraries with relatively large inserts using a pool from a regional library as a composite probe. It is important to note that the target and probe libraries must be based on vectors that share no homology with each other.

MeSH terms

  • Chromosomes, Bacterial / genetics*
  • Chromosomes, Human, Pair 22*
  • Cloning, Molecular / methods*
  • Cosmids / genetics*
  • DNA, Recombinant / genetics
  • Escherichia coli / genetics
  • F Factor / genetics
  • Gene Library*
  • Genome, Human*
  • Humans
  • Nucleic Acid Hybridization

Substances

  • DNA, Recombinant