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Lariat-Dependent Nested PCR for Flanking Sequence Determination

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PCR Protocols

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

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Abstract

Methods detailed in this chapter relate to the use of Lariat-dependent Nested (LaNe) PCR to characterize unknown RNA or DNA sequence flanking known regions. A multitude of approaches designed to determine flanking sequences have been described in the literature. Variously, problems related to these approaches include lack of resolution or failure, depending on experimental context, and complex handling. LaNe-based methods are designed to harness “two-sided” gene-specific PCR with the option of nesting but without the requirement for inefficient and involved enzyme preprocessing steps.

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Acknowledgments

This work was supported by the National Health and Medical Research Council of Australia grant no. 148630 to Professors J.A.M. Graves and M.B. Renfree, Associate Professor V.R. Harley and Dr. A.J. Pask.

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Correspondence to Daniel J. Park .

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© 2011 Humana Press

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Park, D.J. (2011). Lariat-Dependent Nested PCR for Flanking Sequence Determination. In: Park, D. (eds) PCR Protocols. Methods in Molecular Biology, vol 687. Humana Press. https://doi.org/10.1007/978-1-60761-944-4_4

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  • DOI: https://doi.org/10.1007/978-1-60761-944-4_4

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-943-7

  • Online ISBN: 978-1-60761-944-4

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