A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones

Nucleic Acids Res. 1990 May 25;18(10):2887-90. doi: 10.1093/nar/18.10.2887.

Abstract

The recent development of yeast artificial chromosome (YAC) vectors has provided a system for cloning fragments that are over ten times larger than those that can be cloned in more established systems. We have developed a method for the rapid isolation of terminal sequences from YAC clones. The YAC clone is digested with a range of restriction enzymes, a common linker is ligated to the DNA fragments and terminal sequences are amplified using a vector specific primer and a linker specific primer. Sequence data derived from these terminal specific products can be used to design primers for a further round of screening to isolate overlapping clones. The method also provides a convenient method of generating Sequence Tagged Sites for the mapping of complex genomes.

MeSH terms

  • Base Sequence
  • Chromosomes, Fungal
  • Cloning, Molecular
  • DNA Restriction Enzymes
  • Gene Amplification
  • Gene Library*
  • Genetic Techniques*
  • Genetic Vectors
  • Molecular Sequence Data
  • Restriction Mapping
  • Saccharomyces cerevisiae / genetics

Substances

  • DNA Restriction Enzymes

Associated data

  • GENBANK/X52166
  • GENBANK/X52167