Circulating digital PCR method, circulation system, digital PCR chip and preparation method of digital PCR chip

Abstract

The invention discloses a circulating digital polymerase chain reaction (PCR) method, a circulation system, a digital PCR chip and a preparation method of the digital PCR chip. The digital PCR methodincludes the following steps: S1, placing a digital PCR chip subjected to injection processing into a reaction chamber of the circulation system; S2, draining air in the reaction chamber, and pressurizing the circulation system; S3, starting the circulation system for a PCR reaction; S4, depressurizing the circulation system after the PCR reaction is completed; and S5, taking out the digital PCR chip in the reaction chamber, and performing fluorescence signal analysis on the digital PCR chip. The sampling of the chip provided by the invention does not rely on complex equipment such as pumps and valves, and does not need to use high-viscosity thermal polymerization separation oil, the chip does not need to be sealed after the injection is completed, so that the operation is simple; the chiphas a small thickness, fast heat conduction, and a rapid reaction; a constant-temperature liquid storage tank has a large volume and rapid heat exchange, can quickly heat or cool the chip, and has good temperature control, and no temperature overshoot; and the chip has a simple structure and low costs.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection, in particular to a circulation digital PCR method, a circulation system, a digital PCR chip and a preparation method thereof. Background technique [0002] Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments, which can be regarded as special DNA replication outside the body. It has been widely used in molecular biology fields such as gene detection, gene amplification, and genetic engineering, and plays an irreplaceable role in clinical medicine, forensic science, paternity testing, and environmental testing. However, the PCR reaction is exponentially amplified, and can be amplified a million times within tens of minutes, and it is difficult to determine the content of the original PCR template through the PCR product. In order to accurately and quantitatively analyze the nucleic acid content, digital PCR (digital PCR, dPCR) tech...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a circulation type digital PCR method, a circulation system, a digital PCR chip and a preparation method thereof, so as to solve the problems of difficult operation, high cost and long reaction time of the digital PCR method in the prior art

Images