[-癌-] gene expression profiling
作者: biocommander (世翔) 看板: Medicine
標題: [-癌-] gene expression profiling
時間: Fri Apr 23 09:32:43 2010
標題：Validation of a novel, fully integrated and flexible microarray benchtop
facility for gene expression profiling
Received July 6, 2003; Revised September 20, 2003;. Accepted October 9, 2003
Abstract
Here we describe a novel microarray platform that integrates all functions
needed to perform any array-based experiment in a compact instrument on the
researcher’s laboratory benchtop. Oligonucle otide probes are synthesized in
situ via a light- activated process within the channels of a
three-dimensional microfluidic reaction carrier. Arrays can be designed and
produced within hours according to the user’s requirements. They are
processed in a fully automatic workflow. We have characterized this new
platform with regard to dynamic range, discrimination power, reproducibility
and accuracy of biological results. The instrument detects sample RNAs
present at a frequency of 1:100 000. Detection is quantitative over more than
two orders of magnitude. Experiments on four identical arrays with 6398
features each revealed a mean coefficient of variation (CV) value of 0.09 for
the 6398 unprocessed raw intensities indicating high reproducibility. In a
more elaborate experiment targeting 1125 yeast genes from an unbiased
selection, a mean CV of 0.11 on the fold change level was found. Analyzing
the transcriptional response of yeast to osmotic shock, we found that
biological data acquired on our platform are in good agreement with data from
Affymetrix GeneChips, quantitative real-time PCR and—albeit somewhat less
clearly—to data from spotted cDNA arrays obtained from the literature.
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