Selection of appropriate housekeeping genes for quality control

Abstract

The accuracy and validity of gene expression analysis depends on accurate transcript normalization using stably expressed genes. This process compensates for the inherent errors introduced by the measurement process. Putative ‘reference’ or ‘housekeeping genes’ based on their constitutive expression at relatively high levels have traditionally been used for this purpose, although often without any proper validation. More recently, statistical algorithms have now been developed to help validate the suitability of potential reference genes. In this chapter, the concept of normalization in transcript quantification is introduced and a basic protocol for the selection of appropriate reference genes provided.