Normalization methods for analysis of microarray gene-expression data

J Biopharm Stat. 2003 Feb;13(1):57-74. doi: 10.1081/BIP-120017726.

Abstract

This paper investigates subset normalization to adjust for location biases (e.g., splotches) combined with global normalization for intensity biases (e.g., saturation). A data set from a toxicogenomic experiment using the same control and the same treated sample hybridized to six different microarrays is used to contrast the different normalization methods. Simple t-tests were used to compare two samples for dye effects and for treatment effects. The numbers of genes that reproducibly showed significant p-values for the unnormalized data and normalized data from different methods were evaluated for assessment of different normalization methods. The one-sample t-statistic of the ratio of red to green samples was used to test for dye effects using only control data. For treatment effects, in addition to the one-sample t-test of the ratio of the treated to control samples, the two-sample t-test for testing the difference between treated and control samples was also used to compare the two approaches. The method that combines a subset approach (median or lowess fit) for location adjustment with a global lowess fit for intensity adjustment appears to perform well.

Publication types

  • Comparative Study

MeSH terms

  • Bias
  • Data Interpretation, Statistical
  • Fluorescent Dyes
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / statistics & numerical data*
  • Models, Statistical*
  • Normal Distribution
  • Oligonucleotide Array Sequence Analysis / methods
  • Oligonucleotide Array Sequence Analysis / statistics & numerical data*

Substances

  • Fluorescent Dyes