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Novel micro-fluidic chip and digital fluorescent quantitative PCR (Polymerase Chain Reaction) amplification instrument

A microfluidic chip and amplification instrument technology, which is applied in the intersecting field of microfluidic technology and biomedicine, can solve the problems of slow droplet formation speed, small flux, complicated operation, etc., to ensure stability, improve efficiency, The effect of promoting efficiency

Active Publication Date: 2022-04-12
GUANGDONG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of the defects of the prior art, the present invention aims to provide a novel microfluidic chip and digital fluorescent quantitative PCR amplification instrument, using the droplet digital pcr (ddPCR) of the present invention in combination with droplet generation technology and digital PCR technology, It is used to solve the problems of slow droplet formation speed, small throughput, complicated operation and low PCR amplification efficiency commonly existing in the prior art

Method used

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  • Novel micro-fluidic chip and digital fluorescent quantitative PCR (Polymerase Chain Reaction) amplification instrument
  • Novel micro-fluidic chip and digital fluorescent quantitative PCR (Polymerase Chain Reaction) amplification instrument
  • Novel micro-fluidic chip and digital fluorescent quantitative PCR (Polymerase Chain Reaction) amplification instrument

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Embodiment 1

[0044] Chip working principle of the present invention is:

[0045] The sample is injected through the sample port of the chip, and the chip will start to work after each liquid flow is stable. This method uses the pretreatment method of magnetic bead nucleic acid extraction. The premixed liquid mixed with silicone oil, magnetic beads and lysate is injected through the solution inlet of the premixed liquid, and the cell tissue sample liquid inlet is used to inject the Injection of the sample liquid, the two liquid flows are mixed and reacted through the mixing flow channel, so that the cells are lysed, and then the magnetic beads enter to capture nucleic acid molecules; the washing liquid is injected into the eluting solution inlet, and the liquid flow and washing The liquid enters the next mixing flow channel together, reacts and washes away impurities; the first waste liquid collection chamber injects the eluent, the magnetic beads that capture the nucleic acid molecules rea...

Embodiment 2

[0047] After playing the PCR amplification instrument, the opening and closing of the sample compartment and the chip compartment are controlled through the touch screen. After the samples and chips are loaded, they will be aligned and matched through the circuit system and pipeline system, and then the machine will remind the user on the touch screen that the pre-preparation is complete. After being confirmed by the user, the droplet digital PCR microfluidic chip amplification starts. After the droplet is completely generated on the chip, the device will automatically proceed to the next step, start the heating device, and carry out constant temperature amplification at 65°C in the droplet collection chamber of the chip, achieving a 10^9-fold amplification within 30 minutes. After that, PCR fluorescence detection is carried out through the fluorescence detection optical system, and the result will be displayed on the display screen, and the use of the droplet digital PCR devi...

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Abstract

The invention discloses a novel micro-fluidic chip and a digital fluorescent quantitative PCR (Polymerase Chain Reaction) amplification instrument. The chip is formed by combining an upper-layer chip and a lower-layer chip, wherein the upper-layer chip is provided with a solution sample inlet of premixed liquid, a cell tissue sample liquid sample inlet, a washing solution sample inlet, an elution solution sample inlet, a first waste liquid collecting cavity, an oil phase collecting cavity, a deoiling channel and a mixing flow channel; and the lower-layer chip is provided with a multi-channel liquid drop generation flow channel, a second waste liquid collection cavity and a liquid drop collection cavity. The multi-channel liquid drop generation runner is arranged, so that the liquid drop generation efficiency is greatly improved, and the PCR amplification efficiency is remarkably improved. According to the invention, the pretreatment module is combined, and cell lysis is realized by using a magnetic bead method and a nucleic acid extraction method, so that the subsequent PCR reaction is smoothly carried out. And a special deoiling microstructure is arranged, and a passive deoiling method is used, so that the gel is quickly separated from an oil phase after the gel is completed, and the stability of contents is ensured.

Description

technical field [0001] The invention relates to the cross technical field of microfluidic technology and biomedicine, in particular to a novel microfluidic chip and a digital fluorescent quantitative PCR amplification instrument. [0002] technical background [0003] In recent years, as a new type of analysis platform, microfluidic chip technology has the advantages of miniaturization, automation, integration, convenience and speed, and has been widely researched and applied in many fields, such as cell biology, analytical chemistry, environmental Monitoring and protection, forensic identification, drug synthesis and screening, materials science and tissue engineering and other fields. [0004] The microfluidic chip is the main platform for the realization of microfluidic technology, and its main feature is to accommodate the structural units of microfluidics, such as fluid channels, reaction chambers and other functional units. In the research of microfluidic chips, how to...

Claims

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Application Information

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IPC IPC(8): B01L3/00B01L3/02C12N15/10C12Q1/686
CPCY02A50/30
Inventor 杨志达陈雨非宋佳魏鲁明张子榕杨晓丹陈炳钊云涛邓宇
Owner GUANGDONG UNIV OF TECH
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