Cited by 406Gene expression and cancer classificationGenomics and Phylogenetic StudiesOptimal Experimental Design Methods

Statistical Design and Analysis of RNA Sequencing Data

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Auer, Paul L, Doerge, R W·2010·Genetics

Abstract

Next-generation sequencing technologies are quickly becoming the preferred approach for characterizing and quantifying entire genomes. Even though data produced from these technologies are proving to be the most informative of any thus far, very little attention has been paid to fundamental design aspects of data collection and analysis, namely sampling, randomization, replication, and blocking. We discuss these concepts in an RNA sequencing framework. Using simulations we demonstrate the benefits of collecting replicated RNA sequencing data according to well known statistical designs that partition the sources of biological and technical variation. Examples of these designs and their corresponding models are presented with the goal of testing differential expression.

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Crossref · 10.1534/genetics.110.114983

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  • Batch effects can be minimized by appropriate experimental design or removed by batch-correction methods such as COMBAT or ARSyN.

    “These effects can be minimized by appropriate experimental design [51] or, alternatively, removed by batch-correction methods such as COMBAT [52] or ARSyN [20, 53].”

    From A survey of best practices for RNA-seq data analysis (2016) · cites this paper as [51]

  • Without careful experimental design, it is difficult to separate gene expression variability caused by technical artifacts from biological signal.

    “In the absence of careful experimental design, distinguishing gene expression variability due to technical artifacts and biological signal is challenging (Auer and Doerge, 2010).”

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Authors
Auer, P. L., & Doerge, R. W.
Affiliations
Purdue University West Lafayette (US)
Corresponding author
Doerge, R W
Venue
Genetics
Published
2010-06-01
Volume
185
Issue
2
Pages
405-416
ISSN
  • 0016-6731 (ISSN-L)
  • 1943-2631
OpenAlex
W1995070233
Cited by
393
References
53
Citations by year
2026: 5 · 2025: 7 · 2024: 12 · 2023: 11 · 2022: 14 · 2021: 17 · 2020: 24 · 2019: 18 · 2018: 43 · 2017: 25 · 2016: 32 · 2015: 49 · 2014: 42 · 2013: 39 · 2012: 28
FWCI
12.26
Type
journal article
Language
en
Open access
Locations
  • Genetics (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
  • Social Sciences › Decision Sciences › Management Science and Operations Research › Optimal Experimental Design Methods
Keywords
RNA sequencing, statistical design, sampling, randomization, replication, blocking, differential gene expression, biological variation, technical variability, experimental design, gene expression quantification
MeSH terms
Base Sequence, Research Design, Clinical Laboratory Techniques

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