Successful gene expression analysis by multiplex, real-time, one-step RT-PCR, irrespective of the targets amplified

Multiplex, real-time, one-step RT-PCR enables simultaneous quantification of several transcripts in one reaction vessel. This saves time and conserves precious sample material when analyzing the expression of reference and target genes in a particular sample. However, intensive PCR optimization is often required (1). We show that success in multiplex, real-time, one-step RT-PCR can be achieved at the first attempt with the QuantiTect® Multiplex RT-PCR Kit, regardless of the combination of genes analyzed. In each of the multiplex assays carried out, all targets were amplified with high efficiency, which is important for precise relative quantification of gene expression levels by the ΔΔCT method.