Robust RT-qPCR data normalization: validation and selection of internal reference genes during post-experimental data analysis.

ABSTRACT: Reverse transcription and real-time PCR (RT-qPCR) has been widely used for rapid quantification of relative gene expression. To offset technical confounding variations, stably-expressed internal reference genes are measured simultaneously along with target genes for data normalization. Statistic methods have been developed for reference validation; however normalization of RT-qPCR data still remains arbitrary due to pre-experimental determination of particular reference genes. To establish a method for determination of the most stable normalizing factor (NF) across samples for robust data normalization, we measured the expression of 20 candidate reference genes and 7 target genes in 15 Drosophila head cDNA samples using RT-qPCR. The 20 reference genes exhibit sample-specific variation in t

SUBMITTER: Ling D 

PROVIDER: S-EPMC3058000 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature