Selection of chromosome 22-specific clones from human genomic BAC library using a chromosome-specific cosmid library pool.
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.
Kim
U J
UJ
Division of Biology, California Institute of Technology, Pasadena 91125.
Shizuya
H
H
Birren
B
B
Slepak
T
T
de Jong
P
P
Simon
M I
MI
D002876:Chromosomes, Bacterial / Q000235:genetics*; D002892:Chromosomes, Human, Pair 22; D003001:Cloning, Molecular / Q000379:methods*; D003360:Cosmids / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005144:F Factor / Q000235:genetics; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D009693:Nucleic Acid Hybridization
D004274:DNA, Recombinant