Construction and characterization of an eightfold redundant dog genomic bacterial artificial chromosome library

Genomics. 1999 May 15;58(1):9-17. doi: 10.1006/geno.1999.5772.

Abstract

A large insert canine genomic bacterial artificial chromosome (BAC) library was built from a Doberman pinscher. Approximately 166,000 clones were gridded on nine high-density hybridization filters. Insert analysis of randomly selected clones indicated a mean insert size of 155 kb and predicted 8.1 coverage of the canine genome. Two percent of the clones were nonrecombinant. Chromosomal fluorescence in situ hybridization studies of 60 BAC clones indicated no chimerism. The library was hybridized with dog PCR products representing eight genes (ADA, TNFA, GCA, MYB, HOXA, GUSB, THY1, and TOP1). The resulting positive clones were characterized and shown to be compatible with an eightfold redundant library.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Chromosomes, Bacterial
  • Cloning, Molecular
  • DNA / chemistry
  • DNA / genetics
  • DNA Primers
  • Dogs / genetics*
  • Genetic Vectors
  • Genomic Library*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction

Substances

  • DNA Primers
  • DNA

Associated data

  • GENBANK/AF103743
  • GENBANK/AF103744
  • GENBANK/AF103745
  • GENBANK/AF103746
  • GENBANK/AF103747
  • GENBANK/AF103748
  • GENBANK/AF103749
  • GENBANK/AF103750
  • GENBANK/AF103751
  • GENBANK/AF103752
  • GENBANK/AF103753