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Construction of yeast artificial chromosome libraries by pulsed-field gel electrophoresis

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Abstract

Yeast artificial chromosome (YAC) libraries have been constructed from a variety of organisms using different approaches. This protocol outlines in detail the construction of YAC libraries with large inserts using size fractionation of partially digested DNA by pulsed-field gel electrophoresis.

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References

  1. Burke, D. T., Carle, G. F., and Olson, M. V. (1987) Cloning of large DNA segments of exogenous DNA into yeast by means of artificial chromosome vectors.Science 236, 806–812.

    Article  CAS  Google Scholar 

  2. Coulson, A., Waterston, R., Kiff, J., Sulston, J., and Kohara, Y. (1988) Genome linking with yeast artificial chromosomes.Nature 335, 184–186.

    Article  CAS  Google Scholar 

  3. Garza, D., Ajioka, J. W., Burke, D. T., and Haiti, D. L. (1989) Mapping the Drosophila genome with yeast artificial chromosomes.Science 246, 641–646.

    Article  CAS  Google Scholar 

  4. Anand, R., Villasante, A., and Tyler-Smith, C. (1989) Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis.Nucleic Acids Res. 17, 3425–3433.

    Article  CAS  Google Scholar 

  5. McCormick, M. K., Shero, J. H., Cheung, M. C., Kan, Y. W., Hieter, P. A., and Antonarakis, S. E. (1989) Construction of human chromosome 21-specific yeast artificial chromosomes.Proc. Natl. Acad. Sci. USA 86, 9991–9995.

    Article  CAS  Google Scholar 

  6. Albertsen, H. M., Abderrahim, H., Cann, H. C., Dausset, J., Le Paslier, D., and Cohen, D. (1990) Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents.Proc. Natl. Acad. Sci. USA 87, 4256–4260.

    Article  CAS  Google Scholar 

  7. Larin, Z., Monaco, A. P., and Lehrach, H. (1991) Yeast artificial chromosome libraries containing large inserts from mouse and human DNA.Proc. Natl. Acad. Sci. USA 88, 4123–4127.

    Article  CAS  Google Scholar 

  8. Burgers, P. M. J. and Percival, K. J. (1987) Transformation of yeast spheroplasts without cell fusion.Anal. Biochem. 163, 391–397.

    Article  CAS  Google Scholar 

  9. Rothstein, R. (1985) Cloning in yeast, inDNA Cloning, Vol. II (Glover, D. M., ed.), IRL Press, Oxford, pp. 45–65.

    Google Scholar 

  10. Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982)Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

  11. Barlow, D. P. (1992) Preparation, restriction, and hybridization analysis of mammalian genomic DNA for pulse-field gel electrophoresis, in Methods in Molecular Biology, vol 12:Pulse Field Gel Electrophoresis (Burmeister M. and Ulanovsky, L., eds.), Humana, Totowa, NJ, pp. 107–128.

    Google Scholar 

  12. Herrmann, B. G., Barlow, D. P., and Lehrach H. (1987) An inverted duplication of more than 650 Kbp in mouse chromosome 17 mediates unequal but homologous recombination between chromosomes heterozygous for a large inversion.Cell 48, 813–825.

    Article  CAS  Google Scholar 

  13. Hieter, P., Mann, C., Snyder, M., and Davis, R. W. (1985) Mitotic stability of yeast chromosomes: A colony color assay that measures nondisjunction and chromosome loss.Cell 40, 381–392.

    Article  CAS  Google Scholar 

  14. Green, E. D. and Olson, M. V. (1990) Systematic screening of yeast artificial chromosome libraries by use of the polymerase chain reaction.Proc. Natl. Acad. Sci. USA 87, 1213–1217.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Human Genetics Laboratory, Imperial Cancer Research Fund, Institute of Molecular Medicine, John Radcliffe Hospital, 0X3 9DU, Headington, Oxford, UK

    Anthony P. Monaco

  2. Department of Biochemistry, University of Oxford, Oxford, UK

    Zoia Larin

  3. Genome Analysis Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London, UK

    Hans Lehrach

Authors
  1. Anthony P. Monaco
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  2. Zoia Larin
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  3. Hans Lehrach
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Monaco, A.P., Larin, Z. & Lehrach, H. Construction of yeast artificial chromosome libraries by pulsed-field gel electrophoresis. Mol Biotechnol 1, 241–249 (1994). https://doi.org/10.1007/BF02921692

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  • DOI: https://doi.org/10.1007/BF02921692

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