Normalization of cDNA microarray data
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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limma is the only software to allow the use of quantitative weights in loess normalization.
“limma is the only software to allow the use of quantitative weights in loess normalization (38)”
From limma powers differential expression analyses for RNA-sequencing and microarray studies (2015) · cites this paper as [38]
A print-tip loess normalization of the microarray experiments was performed after log-transformation of the background-corrected expression measurements.
“A print-tip loess normalization of the microarray experiments was performed after the log-transformation of the background-corrected expression measurements, as indicated in [67].”
From Adipose tissue transcriptomic signature highlights the pathological relevance of extracellular matrix in human obesity (2008) · cites this paper as [67]
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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
- ORCID
- 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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