Relative quantification

Like many basic research applications, analysis of ocular gene expression is particularly amenable to relative quantification using qPCR (Bustin, 2000; Bustin, 2002; Ginzinger, 2002; Klein, 2002; Walker, 2002) (also discussed in Chapters 2 and 3). This involves calculating changes in gene expression relative to a control population, which should be present in the majority of experimental designs. The control population is totally dependent upon the type of study, and may, for example, be an untreated group (in stimulus or drug-treatment regimes), a specific tissue or cell type (in cross tissue/cell studies), time point zero (in time course studies), wildtype (in transgenic studies) or a specific age group (in developmental or aging studies). Changes are then expressed as a fold change relative to this control population.

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