Digital PCR (polymerase chain reaction) chip and preparation method thereof

Abstract

The invention provides a digital PCR (polymerase chain reaction) chip. The digital PCR chip comprises a droplet production module formed by laminating a cover glass and a glass substrate and a droplet collecting module tightly attached to one surface of the droplet production module, wherein two through holes including a liquid phase sample inlet and an oil phase sample inlet are formed in the cover glass; microflow channels communicated withthe liquid phase sample inlet or the oil phase sample inlet are etched in one surface, attached to the glass substrate, of the cover glass, each microflow channel is divided into two branch channels, and the branch channels pairwise intersect with droplet production ports; the droplet collecting module comprises a cavity with two opening sides, and the droplet production ports are all tightly attached to the inner side of one opening. The digital PCR chip is provided with the independent droplet collecting module, the PCR and the result observation are both performed in the chip, so that the probabilities of pollution and demulsificationare greatly reduced, meanwhile, results of the PCR are observed under a fluorescence microscope in a unified manner, signals reading of droplets is not required to be performed in the chip one by one, and the signal reading time is greatly shortened.

Description

technical field [0001] The invention belongs to the field of nucleic acid detection, and in particular relates to a digital PCR chip and a preparation method thereof. Background technique [0002] Since the polymerase chain reaction (polymerase chain reaction, PCR) was proposed, nucleic acid amplification and quantification technology in vitro has developed into a core technology in the field of molecular biology. At present, PCR-based nucleic acid quantitative detection technology has become the main application technology for early diagnosis of diseases due to its remarkable advantages of fast analysis speed, high sensitivity, high throughput and short diagnosis time, and is widely used in molecular sequencing, gene expression Other research fields such as analysis, gene mutation study and drug screening. [0003] ABI Corporation of the United States has launched real-time fluorescent quantitative PCR (Real-time quantitative PCR, qPCR) technology and related products, whi...

AI Technical Summary

Problems solved by technology

The system can disperse the droplets into millions of droplets. However, the system uses flow analysis technology to analyze millions of droplets one by one in the flow cytometer, and the signal reading takes a long time, resulting in a detection cycle Long, the entire detection process can last up to 3.5 hours

In addition, since the droplets need to be transferred to the flow cytometer for analysis, it is difficult to avoid contamination and demulsification during the transfer process

In addition, when the system generates droplets and collects them for PCR reaction, they are not arranged in a single layer of droplets, but are collected into a sample tank, which will cause some droplets to fuse, resulting in low uniformity of droplets in the system

Images