Normalizing single-cell RNA sequencing data: challenges and opportunities

Nat Methods. 2017 Jun;14(6):565-571. doi: 10.1038/nmeth.4292. Epub 2017 May 15.

Abstract

Single-cell transcriptomics is becoming an important component of the molecular biologist's toolkit. A critical step when analyzing data generated using this technology is normalization. However, normalization is typically performed using methods developed for bulk RNA sequencing or even microarray data, and the suitability of these methods for single-cell transcriptomics has not been assessed. We here discuss commonly used normalization approaches and illustrate how these can produce misleading results. Finally, we present alternative approaches and provide recommendations for single-cell RNA sequencing users.

MeSH terms

  • Algorithms*
  • Data Interpretation, Statistical
  • High-Throughput Nucleotide Sequencing / methods
  • High-Throughput Nucleotide Sequencing / standards*
  • RNA / genetics*
  • Reference Values
  • Sequence Analysis, RNA / standards*
  • Single-Cell Analysis / standards*
  • Transcriptome / genetics*

Substances

  • RNA