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

A large insert canine genomic bacterial artificial chromosome (BAC) library was built from a Doberman pinscher. Approximately 166,000 clones were grid ded on nine high-density hybridization filters. Insert analysis of randomly selected clones indicated a mean insert size of 155 kb and predicted 8.1 c overage of the canine genome. Two percent of the clones were nonrecombinant . Chromosomal fluorescence in situ hybridization studies of 60 BAC clones i ndicated no chimerism. The library was hybridized with dog PCR products rep resenting eight genes (ADA, TNFA, GCA, MYB, HOXA, GUSB, THY1, and TOP1). Th e resulting positive clones were characterized and shown to be compatible w ith an eightfold redundant library. (C) 1999 Academic Press.