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

doi:10.1093/nar/18.10.2887

. 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 Riley¹, R Butler, D Ogilvie, R Finniear, D Jenner, S Powell, R Anand, J C Smith, A F Markham

Affiliations

PMID: 2161516
PMCID: PMC330815
DOI: 10.1093/nar/18.10.2887

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. 1990.

Free PMC article

. 1990 May 25;18(10):2887-90.

doi: 10.1093/nar/18.10.2887.

Authors

J Riley¹, R Butler, D Ogilvie, R Finniear, D Jenner, S Powell, R Anand, J C Smith, A F Markham

Affiliation

¹ ICI Pharmaceuticals, Biotechnology Department, Macclesfield, Cheshire, UK.

PMID: 2161516
PMCID: PMC330815
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.

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References

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[1] Nucleic Acids Res. 1988 Aug 25;16(16):8186 - PubMed

[2] Nucleic Acids Res. 1988 Aug 25;16(16):8186 - PubMed

[3] Proc Natl Acad Sci U S A. 1989 Sep;86(17):6686-90 - PubMed

[4] Proc Natl Acad Sci U S A. 1989 Sep;86(17):6686-90 - PubMed

[5] Nucleic Acids Res. 1988 Sep 12;16(17):8233-43 - PubMed

[6] Nucleic Acids Res. 1988 Sep 12;16(17):8233-43 - PubMed

[7] Science. 1989 Sep 29;245(4925):1434-5 - PubMed

[8] Science. 1989 Sep 29;245(4925):1434-5 - PubMed

[9] Proc Natl Acad Sci U S A. 1986 Oct;83(20):7821-5 - PubMed

[10] Proc Natl Acad Sci U S A. 1986 Oct;83(20):7821-5 - PubMed

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

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

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