Fluorescence pcr refers to the method of adding fluorescent groups to the PCR reaction system, monitoring the whole PCR process in real time by using fluorescence signal accumulation, and finally quantitatively analyzing the unknown template by standard curve. Also known as real-time fluorescence quantitative PCR technology.

The principle of fluorescence PCR technology;

After mixing fluorescein-labeled Taqman probe with template DNA, the thermal cycle of high temperature denaturation, low temperature renaturation and appropriate temperature extension was completed. The Taqman probe complementary to template DNA was cut off and fluorescein was dissociated in the reaction system.

Fluorescence is emitted under the excitation of specific light, and the amplified target gene fragment increases exponentially with the increase of cycle times. The Ct value can be obtained by real-time detection of the corresponding fluorescence signal intensity varying with amplification, and the copy number of the target gene in the sample to be detected can be obtained by using several standards with known template concentrations as controls.