Accurate Real-time Reverse Transcription Quantitative PCR

Within the last few years real-time quantitative PCR has become the method of choice for the accurate quantification of mRNA levels. Compared to previous methods the sensitivity of real-time quantitative PCR improved to the detection limit of up to one single molecule per reaction tube. However, the improved sensitivity leads also to higher demands regarding experimental design. Here we describe an approved protocol to establish mRNA quantification by real-time RT qPCR in a straightforward manner.