Construction and characterization of a human chromosome 2-specific BAC library.
We have constructed a human chromosome 2-specific bacterial artificial chromosome (BAC) library using DNA from the somatic cell hybrid GM10826. The average size of the clones is about 63 kb. The coverage and distribution of the library were estimated by screening with known polymorphic genetic markers and fluorescence in situ hybridization (FISH). Twenty-one markers tested positive when DNA pools prepared from approximately one-sixth of the library were screened with 33 known markers. This is consistent with the theoretical calculation of 63% coverage at one genomic equivalent. This suggested that the coverage of the library is approximately 5-6x. FISH analysis with 54 BACs revealed single site hybridization to chromosome 2, and the clones were distributed randomly on the chromosome. We have also performed direct sequencing of the BAC insert ends to generate sequence-tagged sites suitable for mapping and chromosome walking. This is the first reported human chromosome 2-specific BAC library and should provide a resource for physical mapping and disease searching for this chromosome.
Wang
M
M
Department of Pharmacology, University of North Carolina at Chapel Hill 27599-7365.
Chen
X N
XN
Shouse
S
S
Manson
J
J
Wu
Q
Q
Li
R
R
Wrestler
J
J
Noya
D
D
Sun
Z G
ZG
Korenberg
J
J
D001483:Base Sequence; D002874:Chromosome Mapping; D002889:Chromosomes, Human, Pair 2 / Q000648:ultrastructure; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005819:Genetic Markers; D005821:Genetic Techniques; D006801:Humans; D006822:Hybrid Cells; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D016324:Sequence Tagged Sites
D017931:DNA Primers; D005819:Genetic Markers; D004247:DNA