Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR

Abstract

Until recently expression of housekeeping genes is presumed to remain constant in different cell types, across developmental stages and various experimental conditions hence they are employed as internal controls to normalize gene expression and protein content data. Many studies conducted recently on plant and animal samples have however, reported variable expression of constitutive genes during cellular development 1 and experimental conditions 2-5 . The reason for such variation may be that they are not exclusively involved in performing the basic function of the cell but they also contribute in other cellular processes 6,7 . The variation in their expression is therefore likely and it is advised to consider prior validation of the expression stability of these reference genes, a routine step in all laboratories to accurately normalize the target gene(s) expression. Normalization, a fundamental step is used to compare the target gene expression with the constitutively expressed housekeeping genes. Through this process the technical and biological variations that are introduced during sample preparation or any other step during the experiment are controlled.  doi : 10.5214/ans.0972.7531.210207