Gesamt-Genom-Microarray-Analyse und Target-Validierung mittels qPCR / Whole genome microarray analysis and target validation by using qPCR

Abstract

Abstract In modern pharmaceutical research, microarrays for the analysis of differential gene expression are some of the most valuable tools for the characterization of novel compounds and their biological activity. Moreover, basic clinical research in medicine has benefited tremendously from this rapid development leading to the discovery of new pathophysiological concepts for numerous diseases and to an increased significance for molecular genetic approaches by the characterization of molecular signatures (synonymous for gene expression profile, gene profiling, expression profiling) of diseases or diseased tissues. Extensive data already exist particularly in oncology, where gene profiling has been brought into patient care. For the clinical application of innovative diagnostic procedures, the quest for disease prognosis as well as optimized therapies usually have the highest priority. Particularly in this field, gene expression profiling could help to improve and supplement phenotypic diagnostics and classical laboratory analytics. It is foreseeable that the analysis of a limited number of diagnostically relevant targets and their quantitative measurement by real-time PCR, a reliable technique for this kind of analyses, would suffice. For the identification of diagnostically relevant targets, the research community currently has access to a number of whole-genome arrays or topic-specific arrays, which provide information about the up- or down-regulation of transcripts. These techniques are primarily based upon hybridization of mRNA or cDNA to capture probes and the semi-quantitative measurement of the signal intensity. This article demonstrates the workflow required when setting up a gene expression laboratory for routine analytics in biomedical research and diagnostics.