Accurate mapping of cleavage and polyadenylation sites by 3′ region extraction and deep sequencing

Mainul Hoque, Wencheng Li, Bin Tian

Research output: Chapter in Book/Report/Conference proceedingChapter

19 Scopus citations

Abstract

Deep sequencing of RNA (RNA-seq) is becoming a standard method to study gene expression. While RNA-seq reads cover most regions of an mRNA sequence, they are often depleted in the 3′ end region, making them less amenable for mapping the cleavage and polyadenylation site (pA). A major problem in identifi cation of pA is mispriming at internal A-rich regions and oligo(A) tails when an oligo(dT) primer is used for reverse transcription or sequencing. We recently developed a method named 3′ region extraction and deep sequencing (3′READS), which completely addresses mispriming issues and is straightforward to use. The method accurately maps pAs and allows quantitative analysis of alternative cleavage and polyadenylation (APA) isoforms and gene expression.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages119-129
Number of pages11
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1125
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Cleavage and polyadenylation
  • Deep sequencing
  • Gene expression
  • PolyA site
  • RNA-seq

Fingerprint

Dive into the research topics of 'Accurate mapping of cleavage and polyadenylation sites by 3′ region extraction and deep sequencing'. Together they form a unique fingerprint.

Cite this