Spotting and validation of a genome wide oligonucleotide chip with duplicate measurement of each gene

Biochemical and Biophysical Research Communications
Mads ThomassenT A Kruse

Abstract

The quality of DNA microarray based gene expression data relies on the reproducibility of several steps in a microarray experiment. We have developed a spotted genome wide microarray chip with oligonucleotides printed in duplicate in order to minimise undesirable biases, thereby optimising detection of true differential expression. The validation study design consisted of an assessment of the microarray chip performance using the MessageAmp and FairPlay labelling kits. Intraclass correlation coefficient (ICC) was used to demonstrate that MessageAmp was significantly more reproducible than FairPlay. Further examinations with MessageAmp revealed the applicability of the system. The linear range of the chips was three orders of magnitude, the precision was high, as 95% of measurements deviated less than 1.24-fold from the expected value, and the coefficient of variation for relative expression was 13.6%. Relative quantitation was more reproducible than absolute quantitation and substantial reduction of variance was attained with duplicate spotting. An analysis of variance (ANOVA) demonstrated no significant day-to-day variation.

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Citations

Aug 8, 2014·BioMed Research International·Martin J LarsenTorben A Kruse
Nov 30, 2006·International Journal of Cancer. Journal International Du Cancer·Mads ThomassenTorben A Kruse
Nov 23, 2006·Journal of Microbiological Methods·Patrice FrancoisJacques Schrenzel
Feb 18, 2017·Analytical Chemistry·Josep Ll Acero SánchezCiara K O'Sullivan
Sep 19, 2007·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Mads ThomassenTorben A Kruse

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