Construction of a bovine genomic library of large yeast artificial chromosome clones.
We have constructed a yeast artificial chromosome (YAC) library derived from bull fibroblasts in primary culture. The library consists of 21,500 clones arranged on 224 gridded plates (96 or 8 x 12 positions) and in 2 x 224 96-well microplates maintained as permanent frozen stocks. An average insert size of 750 kb was estimated from the analysis of 200 randomly selected YACs, giving a sixfold coverage of the bovine genome for the total library. A screening strategy based on the polymerase chain reaction that allows the identification of individual clones has been devised. Screening of the library with six locus-specific markers of interest led to the isolation of 34 positive clones. The proportion of chimeric YACs was estimated from the analysis of a subset of the positive clones: 3 of 9 YACs were found to contain noncontiguous DNA.
Libert
F
F
IRIBHN, Brussels, Belgium.
Lefort
A
A
Okimoto
R
R
Womack
J
J
Georges
M
M
D000818:Animals; D002417:Cattle / Q000235:genetics*; D002478:Cells, Cultured; D002874:Chromosome Mapping / Q000662:veterinary; D018244:Chromosomes, Artificial, Yeast; D003001:Cloning, Molecular; D015723:Gene Library; D008297:Male