A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones. |
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| Authors: | J Riley R Butler D Ogilvie R Finniear D Jenner S Powell R Anand J C Smith and A F Markham |
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| Affiliation: | ICI Pharmaceuticals, Biotechnology Department, Macclesfield, Cheshire, UK. |
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| 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. |
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