Simple regression for correcting ΔCt bias in RT-qPCR low-density array data normalization.

ABSTRACT:

Background

Reverse transcription quantitative PCR (RT-qPCR) is considered the gold standard for quantifying relative gene expression. Normalization of RT-qPCR data is commonly achieved by subtracting the Ct values of the internal reference genes from the Ct values of the target genes to obtain ΔCt. ΔCt values are then used to derive ΔΔCt when compared to a control group or to conduct further statistical analysis.

Results

We examined two rheumatoid arthritis RT-qPCR low density array datasets and found that this normalization method introduces substantial bias due to differences in PCR amplification efficiency among genes. This bias results in undesirable correlations between target genes and reference genes, which affect the estimation of fold changes and the tests for diffe

SUBMITTER: Cui X 

PROVIDER: S-EPMC4335788 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature