title: A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones.
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.
authors.affiliation: ICI Pharmaceuticals, Biotechnology Department, Macclesfield, Cheshire, UK.
authors.forename: J
authors.initials: J
authors.lastname: Riley
authors.forename: R
authors.initials: R
authors.lastname: Butler
authors.forename: D
authors.initials: D
authors.lastname: Ogilvie
authors.forename: R
authors.initials: R
authors.lastname: Finniear
authors.forename: D
authors.initials: D
authors.lastname: Jenner
authors.forename: S
authors.initials: S
authors.lastname: Powell
authors.forename: R
authors.initials: R
authors.lastname: Anand
authors.forename: J C
authors.initials: JC
authors.lastname: Smith
authors.forename: A F
authors.initials: AF
authors.lastname: Markham
mesh_terms: D001483:Base Sequence; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D005784:Gene Amplification; D015723:Gene Library; D005821:Genetic Techniques; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D004262:DNA Restriction Enzymes