A simple strand-specific RNA-Seq library preparation protocol combining the Illumina TruSeq RNA and the dUTP methods

Biochem Biophys Res Commun. 2012 Jun 15;422(4):643-6. doi: 10.1016/j.bbrc.2012.05.043. Epub 2012 May 15.


Preserving the original RNA orientation information in RNA-Sequencing (RNA-Seq) experiment is essential to the analysis and understanding of the complexity of mammalian transcriptomes. We describe herein a simple, robust, and time-effective protocol for generating strand-specific RNA-seq libraries suited for the Illumina sequencing platform. We modified the Illumina TruSeq RNA sample preparation by implementing the strand specificity feature using the dUTP method. This protocol uses low amounts of starting material and allows a fast processing within two days. It can be easily implemented and requires only few additional reagents to the original Illumina kit.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Deoxyuracil Nucleotides / chemistry*
  • Deoxyuracil Nucleotides / genetics
  • Gene Expression Profiling / methods*
  • Gene Library*
  • Humans
  • Sequence Analysis, RNA / methods*
  • Transcriptome


  • Deoxyuracil Nucleotides