A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage

Nucleic Acids Res. 1990 Apr 25;18(8):1951-6. doi: 10.1093/nar/18.8.1951.

Abstract

The construction of a yeast artificial chromosome (YAC) primary gridded library of 35,000 clones from human lymphoblastoid (48,XXXX) cell line DNA is described. The average YAC size is approximately 350kb representing a greater than 3.5 times coverage of the genome. The library is stored at -70 degrees C as gridded clones on nylon filters impregnated with 20% glycerol and as glycerol suspensions of individual clones in microtitre plates providing a prolonged multi-user potential. To date we have used 14 single copy probes to screen this library by colony hybridisation as well as PCR and have isolated between 1 and 5 YAC clones for every probe.

MeSH terms

  • Cell Line
  • Chromosome Mapping
  • Chromosomes, Fungal*
  • Cloning, Molecular
  • Gene Library*
  • Humans
  • Restriction Mapping
  • Saccharomyces cerevisiae / genetics
  • Transformation, Genetic