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. 1990 May 25;18(10):2887–2890. 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 1, D Ogilvie 1, R Finniear 1, D Jenner 1, S Powell 1, R Anand 1, J C Smith 1, A F Markham 1
PMCID: PMC330815  PMID: 2161516

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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anand R., Riley J. H., Butler R., Smith J. C., Markham A. F. A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage. Nucleic Acids Res. 1990 Apr 25;18(8):1951–1956. doi: 10.1093/nar/18.8.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anand R., Villasante A., Tyler-Smith C. Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis. Nucleic Acids Res. 1989 May 11;17(9):3425–3433. doi: 10.1093/nar/17.9.3425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brownstein B. H., Silverman G. A., Little R. D., Burke D. T., Korsmeyer S. J., Schlessinger D., Olson M. V. Isolation of single-copy human genes from a library of yeast artificial chromosome clones. Science. 1989 Jun 16;244(4910):1348–1351. doi: 10.1126/science.2544027. [DOI] [PubMed] [Google Scholar]
  4. Burke D. T., Carle G. F., Olson M. V. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science. 1987 May 15;236(4803):806–812. doi: 10.1126/science.3033825. [DOI] [PubMed] [Google Scholar]
  5. Coulson A., Sulston J., Brenner S., Karn J. Toward a physical map of the genome of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7821–7825. doi: 10.1073/pnas.83.20.7821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coulson A., Waterston R., Kiff J., Sulston J., Kohara Y. Genome linking with yeast artificial chromosomes. Nature. 1988 Sep 8;335(6186):184–186. doi: 10.1038/335184a0. [DOI] [PubMed] [Google Scholar]
  7. Nelson D. L., Ledbetter S. A., Corbo L., Victoria M. F., Ramírez-Solis R., Webster T. D., Ledbetter D. H., Caskey C. T. Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6686–6690. doi: 10.1073/pnas.86.17.6686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Newton C. R., Kalsheker N., Graham A., Powell S., Gammack A., Riley J., Markham A. F. Diagnosis of alpha 1-antitrypsin deficiency by enzymatic amplification of human genomic DNA and direct sequencing of polymerase chain reaction products. Nucleic Acids Res. 1988 Sep 12;16(17):8233–8243. doi: 10.1093/nar/16.17.8233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ochman H., Gerber A. S., Hartl D. L. Genetic applications of an inverse polymerase chain reaction. Genetics. 1988 Nov;120(3):621–623. doi: 10.1093/genetics/120.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Olson M., Hood L., Cantor C., Botstein D. A common language for physical mapping of the human genome. Science. 1989 Sep 29;245(4925):1434–1435. doi: 10.1126/science.2781285. [DOI] [PubMed] [Google Scholar]
  11. Silverman G. A., Ye R. D., Pollock K. M., Sadler J. E., Korsmeyer S. J. Use of yeast artificial chromosome clones for mapping and walking within human chromosome segment 18q21.3. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7485–7489. doi: 10.1073/pnas.86.19.7485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Traver C. N., Klapholz S., Hyman R. W., Davis R. W. Rapid screening of a human genomic library in yeast artificial chromosomes for single-copy sequences. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5898–5902. doi: 10.1073/pnas.86.15.5898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Triglia T., Peterson M. G., Kemp D. J. A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences. Nucleic Acids Res. 1988 Aug 25;16(16):8186–8186. doi: 10.1093/nar/16.16.8186. [DOI] [PMC free article] [PubMed] [Google Scholar]

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