Fluorescence-based quantitative analysis for mRNA in RT-PCR process

A DNA fragment that forms the template of a specific mRNA can be synthesized by reverse transcription PCR, In due number of cycle, the concentration of PCR product increases proportionally to that of the template cDNA, i. e., the corresponding mRNA in RT. Since the fluorescence quantam yield of ethidium bromide greatly increased when it was inserted into the DNA double helix, the extent of expression of mRNA can be calculated by determining the fluorescence intensity under selected excitation and emission wavelengths of PCR cDNA after a proper number of PCR cycle.