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Analysis of Yeast Artificial Chromosome DNA by Restriction Digestion, Southern Blotting Nucleic Acid Hybridization, and Polymerase Chain Reaction

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YAC Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 349))

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Abstract

Restriction enzyme analysis of yeast artificial chromosome (YAC) clones in combination with Southern blotting and polymerase chain reaction (PCR) is necessary to establish the integrity of the DNA contained within the YAC clone, as well as obtain information on the integrity of the cloned DNA. Restriction analysis is also a useful tool that allows a direct comparison between endogenous genomic DNA and the DNA fragment present within a YAC clone. This chapter summarizes the techniques of enzyme digestion of YAC DNA and the separation of the DNA fragments by pulse-field gel electrophoresis. The protocols of Southern blotting and PCR used to identify the cloned DNA fragments will also be described.

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References

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Author information

Authors and Affiliations

  1. Department of Physiology, White Block, University of Liverpool, Crown Street, Liverpool, England

    Sylvia Vasiliou & John Peter Quinn

Authors
  1. Sylvia Vasiliou
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  2. John Peter Quinn
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Editor information

Editors and Affiliations

  1. School of Medical Sciences, College of Life Sciences and Medicine, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen, Scotland

    Alasdair MacKenzie

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© 2006 Humana Press Inc., Totowa NJ

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Vasiliou, S., Quinn, J.P. (2006). Analysis of Yeast Artificial Chromosome DNA by Restriction Digestion, Southern Blotting Nucleic Acid Hybridization, and Polymerase Chain Reaction. In: MacKenzie, A. (eds) YAC Protocols. Methods in Molecular Biology™, vol 349. Humana Press. https://doi.org/10.1385/1-59745-158-4:33

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  • DOI: https://doi.org/10.1385/1-59745-158-4:33

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-612-2

  • Online ISBN: 978-1-59745-158-1

  • eBook Packages: Springer Protocols

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