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

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

J Riley  1 , R Butler, D Ogilvie, R Finniear, D Jenner, S Powell, R Anand, J C Smith, A F Markham
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Free PMC article

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

J Riley et al. Nucleic Acids Res. .
Free PMC article

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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References

    1. Nucleic Acids Res. 1988 Aug 25;16(16):8186 - PubMed
    1. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6686-90 - PubMed
    1. Nucleic Acids Res. 1988 Sep 12;16(17):8233-43 - PubMed
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