Detection and quantitative estimation of spurious double stranded DNA formation during reverse transcription in bacteria using tagRNA-seq

RNA Biol. 2015;12(9):1067-9. doi: 10.1080/15476286.2015.1071010.

Abstract

Standard RNA-seq has a well know tendency to generate "ghost" antisense reads due to formation of spurious second strand cDNA in the sequencing process. We recently reported on a novel variant of RNA-seq coined "tagRNA-seq" introduced for the purpose of distinguishing primary from processed transcripts in bacteria. Incidentally, the additional information provided by the tags is also very suitable for detection of true anti-sense RNA transcripts and quantification of spurious antisense signals in a sample. We briefly explain how to perform such a detection and illustrate on previously published datasets.

Keywords: DNA; antisense RNA; complementary; spurious second strand cDNA; tagRNA-seq; transcript discovery; transcriptome.

Publication types

  • Letter

MeSH terms

  • Bacteria / genetics*
  • DNA / genetics*
  • High-Throughput Nucleotide Sequencing
  • Reverse Transcription*
  • Sequence Analysis, RNA

Substances

  • DNA