A minimum variance method for genome-wide data-driven normalization of quantitative real-time polymerase chain reaction expression data

Advances in multiplex qRT-PCR have enabled increasingly accurate and robust quantification of RNA, even at lower concentrations, facilitating RNA expression profiling in clinical and environmental samples. Here we describe a data-driven qRT-PCR normalization method, the minimum variance method, and evaluate it on clinically derived Mycobacterium tuberculosis samples with variable transcript detection percentages. For moderate to significant amounts of nondetection (similar to 50%), our minimum variance method consistently produces the lowest false discovery rates compared to commonly used data-driven normalization methods. (C) 2014 The Authors. Published by Elsevier Inc.