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Isolation of Genomic Insertion Sites of Proviruses Using Splinkerette-PCR-Based Procedures

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PCR Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 687))

Abstract

The availability of whole genomic sequences provides a great framework for biologists to address a broad range of scientific questions. However, functions of most mammalian genes remain obscure. The forward genetics strategy of insertional mutagenesis uses DNA mutagens such as retroviruses and transposable elements; this strategy represents a powerful approach to functional genomics. A variety of methods to uncover insertion sites have been described. This chapter details SplinkTA-PCR and SplinkBlunt-PCR, modified from splinkerette-PCR, for mapping chromosomally the insertion sites of a murine leukemia virus that causes leukemia in the BXH-2 strain of mice. These protocols are easy to use, reliable, and efficient.

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Acknowledgments

The author would like to express thanks to Ms. Marianna L. Wong and Mr. John W. Myers III for their help with grammatical editing. This work is supported by the National Basic Research Program of China (973 Program, No. 2011CB933501, to Dr. Bin Yin) and the NSFC Project (No. 81070417, to Dr. Bin Yin).

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Authors and Affiliations

  1. Cyrus Tang Hematology Center, Jiangsu Institute of Hematology, First Affiliated Hospital, Soochow University, Suzhou, China

    Bin Yin

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  1. Bin Yin
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Editors and Affiliations

  1. , Department of Pathology, The University of Melbourne, Corner of Grattan St & Royal Parade, Parkville, 3010, Victoria, Australia

    Daniel J. Park

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© 2011 Humana Press

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Yin, B. (2011). Isolation of Genomic Insertion Sites of Proviruses Using Splinkerette-PCR-Based Procedures. In: Park, D. (eds) PCR Protocols. Methods in Molecular Biology, vol 687. Humana Press. https://doi.org/10.1007/978-1-60761-944-4_3

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  • DOI: https://doi.org/10.1007/978-1-60761-944-4_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-943-7

  • Online ISBN: 978-1-60761-944-4

  • eBook Packages: Springer Protocols

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