Method and device for carrying out polymerase chain reaction under constant-temperature heat source

Abstract

The invention provides a method and device for carrying out polymerase chain reaction under a constant-temperature heat source, and specifically relates to a method for providing heat to or taking away heat from a specific area of a reaction test tube based on a Rayleigh-Benard theory to build a temperature gradient from bottom to top of reagents in the reaction tube, spontaneously carrying out convection under the condition that the reaction reagents are unevenly heated, and carrying out corresponding PCR (Polymerase Chain Reaction) amplification when the reaction reagents flow through different temperature areas. The invention also discloses a device for carrying out the reaction of the method and in-time fluorescence detection.

Description

technical field [0001] The invention relates to a polymerase chain reaction method and device. More specifically, the present invention relates to the principle of establishing a bottom-up temperature gradient of a liquid based on heat convection, a method for spontaneous convection when the liquid is heated, and corresponding PCR amplification when it flows through different temperature regions, and a corresponding device. Background technique: [0002] Polymerase chain reaction technology (hereinafter referred to as PCR technology) is a technology for rapidly amplifying DNA in vitro. Each cycle includes three processes of denaturation, annealing and extension. After each cycle, the number of target nucleic acid molecules is doubled. , after 30-40 cycles, the number of target nucleic acid molecules was amplified to nearly 10 9 PCR is a method to obtain a large number of target DNA fragments in vitro, which is convenient for further analysis and inspection of nucleic acid m...

AI Technical Summary

Problems solved by technology

The defect of this device is: the reaction volume is large, that is, the system usually has a large volume and heat capacity, and it generally takes 2-3 hours for conventional PCR to complete 30 cycles, most of which time is consumed in the heating and cooling process, that is, the metal The block reaches the equilibrium temperature and transfers heat to the PCR reaction solution through the reaction tube, so it is difficult for PCR to achieve high efficiency and high throughput

However, there are still defects in this method: firstly, the reagent needs to fill the entire reaction chamber, which needs to be sealed, and there is a potential leakage problem; secondly, the reagent is directly injected into the reaction chamber, resulting in direct contact between the heater and the reagent, and there is a potential pollution problem

[0008] Therefore, there is an urgent need for a new polymerase chain reaction amplification method and corresponding devices in the prior art to solve the problems of easy pollution and unstable reaction.

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