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https://hdl.handle.net/2440/24067
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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: the international journal for all aspects of chromosome and nuclear biology, 2006; 14(6):657-664 |
Publisher: | Kluwer Academic Publ |
Issue Date: | 2006 |
ISSN: | 0967-3849 1573-6849 |
Statement of Responsibility: | 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 |
DOI: | 10.1007/s10577-006-1076-z |
Published version: | http://dx.doi.org/10.1007/s10577-006-1076-z |
Appears in Collections: | Aurora harvest 2 Environment Institute Leaders publications Molecular and Biomedical Science publications |
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