Chromosome Research

, Volume 14, Issue 6, pp 657–664 | Cite as

Construction of a highly enriched marsupial Y chromosome-specific BAC sub-library using isolated Y chromosomes

  • N. Sankovic
  • M. L. Delbridge
  • F. Grützner
  • M. A. Ferguson-Smith
  • P. C. M. O’Brien
  • J. A. Marshall Graves
Article

Abstract

The Y chromosome is perhaps the most interesting element of the mammalian genome but comparative analysis of the Y chromosome has been impeded by the difficulty of assembling a shotgun sequence of the Y. BAC-based sequencing has been successful for the human and chimpanzee Y but is difficult to do efficiently for an atypical mammalian model species (Skaletsky et al.2003, Kuroki et al.2006). We show how Y-specific sub-libraries can be efficiently constructed using DNA amplified from microdissected or flow-sorted Y chromosomes. A Bacterial Artificial Chromosome (BAC) library was constructed from the model marsupial, the tammar wallaby (Macropus eugenii). We screened this library for Y chromosome-derived BAC clones using DNA from both a microdissected Y chromosome and a flow-sorted Y chromosome in order to create a Y chromosome-specific sub-library. We expected that the tammar wallaby Y chromosome should detect ∼100 clones from the 2.2 times redundant library. The microdissected Y DNA detected 85 clones, 82% of which mapped to the Y chromosome and the flow-sorted Y DNA detected 71 clones, 48% of which mapped to the Y chromosome. Overall, this represented a ∼330-fold enrichment for Y chromosome clones. This presents an ideal method for the creation of highly enriched chromosome-specific sub-libraries suitable for BAC-based sequencing of the Y chromosome of any mammalian species.

Key words

BAC library Macropus eugenii tammar wallaby Y chromosome 

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Copyright information

© Springer 2006

Authors and Affiliations

  • N. Sankovic
    • 1
    • 2
  • M. L. Delbridge
    • 1
  • F. Grützner
    • 1
    • 3
  • M. A. Ferguson-Smith
    • 4
  • P. C. M. O’Brien
    • 4
  • J. A. Marshall Graves
    • 1
  1. 1.Comparative Genomics Group, Research School of Biological SciencesAustralian National UniversityCanberraAustralia
  2. 2.Department of ZoologyUniversity of MelbourneParkvilleAustralia
  3. 3.School of Molecular and Biomedical ScienceUniversity of AdelaideAdelaideAustralia
  4. 4.Department of Veterinary MedicineCambridge UniversityCambridgeUK

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