RT-qPCR stands for quantitative reverse transcription polymerase chain reaction, the “RT” not to be mistaken for “real time”. Unlike qPCR that uses a DNA template, the starting material for RT-qPCR is RNA. Therefore, protocols incorporate a reverse transcription step to convert the RNA to cDNA before the normal qPCR amplification process commences. This may be done all in one reaction tube (one-step) or sequentially with reverse transcription taking place in a separate reaction to the qPCR amplification (two-step).