Cited by 1,871Gene expression and cancer classificationGenomics and Phylogenetic StudiesBioinformatics and Genomic Networks

Normalization of cDNA microarray data

Smyth, Gordon K, Speed, Terry·2003·Methods

Abstract

Normalization means to adjust microarray data for effects which arise from variation in the technology rather than from biological differences between the RNA samples or between the printed probes. This paper describes normalization methods based on the fact that dye balance typically varies with spot intensity and with spatial position on the array. Print-tip loess normalization provides a well-tested general purpose normalization method which has given good results on a wide range of arrays. The method may be refined by using quality weights for individual spots. The method is best combined with diagnostic plots of the data which display the spatial and intensity trends. When diagnostic plots show that biases still remain in the data after normalization, further normalization steps such as plate-order normalization or scale-normalization between the arrays may be undertaken. Composite normalization may be used when control spots are available which are known to be not differentially expressed. Variations on loess normalization include global loess normalization and two-dimensional normalization. Detailed commands are given to implement the normalization techniques using freely available software.

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Crossref · 10.1016/s1046-2023(03)00155-5

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Authors
Smyth, G. K., & Speed, T.
Affiliations
Walter and Eliza Hall Institute of Medical Research (AU)
Corresponding author
Smyth, Gordon K
Venue
Methods
Publisher
Elsevier BV
Published
2003-12
Volume
31
Issue
4
Pages
265-273
ISSN
1046-2023 (ISSN-L)
OpenAlex
W2013444425
Cited by
1,871
References
8
Citations by year
2026: 6 · 2025: 14 · 2024: 26 · 2023: 38 · 2022: 35 · 2021: 45 · 2020: 47 · 2019: 54 · 2018: 43 · 2017: 68 · 2016: 73 · 2015: 104 · 2014: 130 · 2013: 145 · 2012: 165
FWCI
17.58
Type
journal article
Language
en
Open access
closed
Locations
  • Methods (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institute on Aging
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
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bioinformatics and Genomic Networks
Keywords
scale normalization, differential gene expression
MeSH terms
Reference Values, Software, DNA, Complementary, Oligonucleotide Array Sequence Analysis

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