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Characterizing cDNA Ends by Circular RACE

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Molecular Methods for Evolutionary Genetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 772))

Abstract

Rapid amplification of cDNA ends (RACE) is a widely used PCR-based method to identify the 5′ and 3′ ends of cDNA transcripts from partial cDNAs. While conceptually simple, this method often requires substantial optimization before accurate end identification is achieved. This is due in part to the anchoring of a universal primer to a cDNA or mRNA for PCR, which can lead to the generation of nonspecific amplification. Here, we describe an improvement of the original RACE method, circular RACE, which can be used to simultaneously identify both the 5′ and 3′ end of a target cDNA.

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Acknowledgment

We thank Andres Bendesky for critical reading of this protocol.

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Correspondence to Patrick T. McGrath .

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© 2012 Springer Science+Business Media, LLC

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McGrath, P.T. (2012). Characterizing cDNA Ends by Circular RACE. In: Orgogozo, V., Rockman, M. (eds) Molecular Methods for Evolutionary Genetics. Methods in Molecular Biology, vol 772. Humana Press. https://doi.org/10.1007/978-1-61779-228-1_15

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  • DOI: https://doi.org/10.1007/978-1-61779-228-1_15

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-227-4

  • Online ISBN: 978-1-61779-228-1

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