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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.

SUBMITTER: Riley J

PROVIDER: S-EPMC330815 | biostudies-other | 1990 May

REPOSITORIES: biostudies-other

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

Riley J J Butler R R Ogilvie D D Finniear R R Jenner D D Powell S S Anand R R Smith J C JC Markham A F AF

Nucleic acids research 19900501 10

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. ...[more]

PMID: 2161516

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