GB/T 36433-2018Textiles—DNA quantitative analysis of cashmere and wool mixture—Fluorescence PCR method (English PDF)

3.1 Polymerase chain reaction; PCR

Under the catalysis of DNA polymerase, using the parent strand DNA as a template and the specific primers as the starting points, 4 kinds of single nucleotides (dNTP) are The substrate, through a denaturing, annealing, and extended cyclic reaction, causes a very small amount of specific fragments of DNA to be amplified millions of times in a matter of hours. In vitro amplification of DNA by chain reaction.

3.2 Real-time fluorescence PCR

Fluorescence groups are added to the PCR reaction system to monitor the entire PCR process in real-time using fluorescence signal accumulation and then pass the curve to unknown Template for quantitative or qualitative analysis of DNA amplification methods.

3.3 Primer

In the nucleic acid synthesis reaction, a small piece of single-stranded DNA that functions as a starting point for the extension of each polynucleotide strand and acts Or RNA.

3.4 TaqMan probe

One fluorophore is attached to the 5' end of the probe and the quencher is the 3' end of the oligonucleotide.

Note: A specific fluorescent probe is added at the same time that a pair of primers is added during PCR amplification. When the probe is intact, the fluorescence signal emitted by the reporter group is quenched.

The group absorbs; during PCR amplification, the Taq enzyme's 5'-3' exonuclease activity degrades the probe, causing the reporter and quencher fluorophores to separate.

The fluorescence monitoring system can receive the fluorescence signal, that is, one fluorescent molecule is formed for each DNA strand to be amplified, and the fluorescence signal is accumulated.