A 3' RACE protocol to confirm polyadenylation sites

Liuyin Ma, Arthur G. Hunt

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

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.

Original languageEnglish
Title of host publicationPolyadenylation in Plants
Subtitle of host publicationMethods and Protocols
Pages135-144
Number of pages10
ISBN (Electronic)9781493921751
DOIs
StatePublished - Dec 8 2014

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media New York 2015. All rights are reserved.

Keywords

  • 3' RACE protocol
  • Ion torrent
  • MiSeq
  • Polyadenylation sites

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (all)
  • Medicine (all)
  • Biochemistry, Genetics and Molecular Biology (all)

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