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Type: Journal article
Title: Construction of a highly enriched marsupial Y chromosome-specific BAC sub-library using isolated Y chromosomes
Author: Sankovic, N.
Delbridge, M.
Grutzner, F.
Ferguson-Smith, M.
O'Brien, P.
Graves, J.
Citation: Chromosome Research, 2006; 14(6):657-664
Publisher: Kluwer Academic Publ
Issue Date: 2006
ISSN: 0967-3849
Statement of
N. Sankovic, M. L. Delbridge, F. Grützner, M. A. Ferguson-Smith, P. C. M. O’Brien and J. A. Marshall Graves
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 approximately 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 approximately 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.
Keywords: BAC library; Macropus eugenii; tammar wallaby; Y chromosome
Description: © Springer 2006
RMID: 0020061261
DOI: 10.1007/s10577-006-1076-z
Appears in Collections:Molecular and Biomedical Science publications
Environment Institute Leaders publications

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