a Whole Genome Array Approach - Jacobs University

Background
coupling of fluorescent dyes (Beier et al. 2004). Equal amounts of the labelled target cDNA
(reference + sample) are combined, co-hybridised under stringent conditions onto a
microarray replicate and washed in a defined procedure. Afterwards the slide is scanned using
a specialised laser scanner to record the emission of the two fluorochromes for each spot on
the microarray after excitation at a given wavelength. The intensity of the fluorescence
emission signals on each spot is proportional to transcript levels in the biological samples and
the fluorescent signal from each gene probe is quantified by using specialised image
processing software. The output can then be analysed and normalised using different analysis
software. A standard method to evaluate the results is quantitative Real-Time (RT)-PCR.
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