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A 3′ RACE Protocol to Confirm Polyadenylation Sites

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Polyadenylation in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1255))

Abstract

A routine procedure in the study of polyadenylation involves the determinations of the junctions of the mRNA body and the poly(A) tail (the poly(A) site). This is typically accomplished by selectively amplifying the 3′-ends of cDNAs (3′-Rapid Amplification of cDNA Ends, or 3′-RACE), followed by sequencing of individual clones. We have developed a modification of the standard 3′-RACE protocol that couples high specificity with the ability to characterize thousands (or more) of individual sequences. This protocol may be used for numerous purposes, including the confirmation of results obtained using high throughput sequencing technologies.

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Acknowledgements

This work was supported by US National Science Foundation (IOS-081781). In addition, Liuyin Ma was a recipient of a scholarship from the China Scholarship Council.

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Authors and Affiliations

  1. College of the Environment and Ecology, Xiamen University, Xiamen, 361102, China

    Liuyin Ma

  2. Department of Plant and Soil Sciences, University of Kentucky, 105 Plant Science Building, 1405 Veterans Drive, Lexington, KY, 40546, USA

    Arthur G. Hunt

Authors
  1. Liuyin Ma
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  2. Arthur G. Hunt
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Corresponding author

Correspondence to Arthur G. Hunt .

Editor information

Editors and Affiliations

  1. Department of Plant and Soil Sciences, University of Kentucky, Lexington, Kentucky, USA

    Arthur G. Hunt

  2. Department of Biology, Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, College of the Environment and Ecology, Miami University, Oxford, OH, USA and Xiamen University, Xiamen, Fujian, Fujian, China

    Qingshun Quinn Li

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Ma, L., Hunt, A.G. (2015). A 3′ RACE Protocol to Confirm Polyadenylation Sites. In: Hunt, A., Li, Q. (eds) Polyadenylation in Plants. Methods in Molecular Biology, vol 1255. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2175-1_12

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  • DOI: https://doi.org/10.1007/978-1-4939-2175-1_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2174-4

  • Online ISBN: 978-1-4939-2175-1

  • eBook Packages: Springer Protocols

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