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YAC Library Screening

Hybridization and PCR-Based Screening Protocols
  • Charlotte G. Cole
  • John E. Collins
  • Ian Dunham
Protocol
  • 85 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Yeast artificial chromosome (YAC) libraries stored in microtiter plates are available for screening as either complex PCR pools or hybridization filters generated from YACs gridded at high densities (see  Chapter 57). Different libraries may be available as either PCR pools, hybridization filters, or both. Consequently, screening strategies have been designed that rely solely on either technique, or use a combined approach. Clearly, access to the YAC library microtiter plates and an automatic gridding system allows the user greater flexibility and is an advantage in large mapping projects.

Keywords

Microtiter Plate Hybridization Buffer Yeast Artificial Chromosome Primary Pool Saran Wrap 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Bentley, D. R., Todd, C., Collins J., Holland, J., Dunham, I., Hassock, S., et al. (1992) The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries. Genomics 12, 534–541.PubMedCrossRefGoogle Scholar
  2. 2.
    McKeown, G., Watson, A., Karunaratne, K., and Bentley D. (1993) High throughput filter preparation robot. Genome Sci. Technol. (special first issue), Program and Abstracts Genome Sequencing and Analysis Conference V, October 23–27, 1993, p. 56 (abstract C 21).Google Scholar
  3. 3.
    Green, E. D. and Olson, M. V. (1990) Systematic screening of yeast artificial-chromosome libraries using the polymerase chain reaction. Proc. Natl. Acad. Sci. USA 87, 1213–1217.PubMedCrossRefGoogle Scholar
  4. 4.
    Amemiya, C. T., Alegria-Hartman, M. J., Aslanidis, C., Chen, C., Nikolic, J., Gringrich, J. C., and De Jong, P. J. (1992) A two-dimensional YAC pooling strategy for library screening via STS and Alu-PCR methods. Nucleic Acids Res. 20, 2559–2563.PubMedCrossRefGoogle Scholar
  5. 5.
    Chumakov, I., Rigault, P., Guillou, S., Ougen, P., Billaut, A., Guasconi, G., et al. (1992) Continuum of overlapping clones spanning the entire human chromosome 21q. Nature (Lond.) 359, 380–387.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Charlotte G. Cole
    • 1
  • John E. Collins
    • 1
  • Ian Dunham
    • 1
  1. 1.The Sanger CentreWellcome Trust Genome CampusCambridgeUK

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