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

Mads Thomassen*, Vibe Skov, Freyja Eiriksdottir, Qihua Tan, Kirsten Jochumsen, Niels Fritzner, Klaus Brusgaard, Jesper Dahlgaard, Torben A. Kruse

*Corresponding author af dette arbejde

    Publikation: Bidrag til tidsskriftArtikelForskningpeer review

    Abstrakt

    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.

    OriginalsprogEngelsk
    Sider (fra-til)1111-1120
    Antal sider10
    TidsskriftBiochemical and Biophysical Research Communications
    Vol/bind344
    Udgave nummer4
    DOI
    StatusUdgivet - 16 jun. 2006

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