Digital PCR detection method

Abstract

The invention relates to a digital PCR detection method, when the nucleic acid amplification reaction liquid to be detected is prepared, genotyping, mutation scanning, methylation research and the like can be realized by using dyes, high resolution and sensitivity are achieved, and the detection cost is reduced. Moreover, a polymerase chain reaction of a micro-droplet array can be completed on a same highly-integrated digital PCR detector, and melting curve analysis is carried out on a PCR product after PCR amplification of the micro-droplet array. Meanwhile, through the digital PCR detectionmethod, a fluorescence curve and a melting curve of the micro-droplet array can be obtained, and traceless connection between real-time monitoring of the whole PCR amplification process and melting curve analysis of the PCR product can be completely achieved. Therefore, copy number identification based on different fluorescence curves, identification of nucleic acid amplification characteristics based on different characteristic melting curves and typing, mutation scanning and the like of genes through high-resolution melting curves are realized, and digital PCR detection is completed more comprehensively, simply, conveniently and efficiently.

Description

technical field [0001] The present application relates to the field of digital PCR detection and analysis, in particular to a digital PCR detection method. Background technique [0002] In recent years, digital PCR (Digital PCR, dPCR) technology has developed rapidly. Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. Digital PCR includes droplet PCR detection method and chip detection method. In the chip-based detection method, the number of effective reaction chambers on a single chip is generally only a few thousand, which is far less than that of the micro-droplet method. The dynamic range of the chip-based digital PCR is narrower than that of the micro-droplet method. [0003] The droplet digital PCR system performs droplet treatment on the sample before the traditional PCR amplification, that is, the reaction system containing nucleic acid molecules is divided into tens of thousands of nanoliter droplets, each of ...

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Problems solved by technology

[0004] Based on this, it is necessary to provide a simple and efficient digital PCR detection method for the problems of heavy workload, time-consuming and low efficiency of the traditional digital PCR detection method

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