Please use this identifier to cite or link to this item:
|Scopus||Web of Science®||Altmetric|
|Title:||Construction of a highly enriched marsupial Y chromosome-specific BAC sub-library using isolated Y chromosomes|
|Citation:||Chromosome Research, 2006; 14(6):657-664|
|Publisher:||Kluwer Academic Publ|
|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|
|Appears in Collections:||Molecular and Biomedical Science publications|
Environment Institute Leaders publications
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.