Enzyme- and gene-specific biases in reverse transcription of RNA raise concerns for evaluating gene expression

Sci Rep. 2020 May 18;10(1):8151. doi: 10.1038/s41598-020-65005-0.

Abstract

Reverse transcription is the first step of most analyses of gene expression, yet the quantitative biases it introduces are largely overlooked. Following a series of purpose-designed systematic experiments we cherry-pick examples of various biases introduced by reverse transcription, and alert the "gene expression community" to the pitfalls and improved practice of this fundamental technique.

MeSH terms

  • Cell Line, Tumor
  • Gene Expression Profiling
  • Humans
  • RNA / genetics*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcription*

Substances

  • RNA