Digital PCR (Polymerase Chain Reaction) method of integrated micro-droplet chip
A droplet, digital technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of low integration and automation
Pending Publication Date: 2022-03-18
TARGETINGONE TECH (BEIJING) CORP
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Problems solved by technology
[0005] Therefore, the technical problem to be solved by the present invention is to provide a digital PCR method for an integrated micro-droplet chip, so as to overcome the shortcomings in the prior art that droplet generation and detection are completed in different droplet chips, and the degree of integration and automation is low.
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[0031] see in conjunction Figure 1 to Figure 9 As shown, according to the embodiment of the present invention, an integrated micro-droplet chip is provided, including a chip body 1, the chip body 1 has a reaction chamber 11 and a sample loading chamber 12, and the chip body 1 is configured with a micro-fluid Droplet generation structure, oil-liquid interface 31, gas-liquid interface 32 and fluorescence detection area 33, the gas-liquid interface 32 communicates with the reaction chamber 11, the sample adding chamber 12 communicates with the micro-droplet generation structure, the The oil interface 31 is in communication with the micro droplet generating structure;
[0032] When the droplets are generated, a first pressure difference is formed between the sample adding chamber 12 and the gas-liquid interface 32, a second pressure difference is formed between the oil-liquid interface 31 and the gas-liquid interface 32, and the first The first pressure difference and the second...
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The invention provides a digital PCR (Polymerase Chain Reaction) method of an integrated micro-droplet chip, which comprises the following steps: controlling a first pressure difference between a sample adding cavity and a gas-liquid interface and a second pressure difference between an oil interface and the gas-liquid interface to respectively drive a sample in the sample adding cavity and generated oil of the oil interface to enter a micro-droplet generating structure, the generated micro-droplets are stored in the reaction cavity; placing the reaction cavity in a heating module for heating amplification; external pressure is controlled to drive detection pushing oil to enter the reaction cavity from the gas-liquid connector, so that micro-droplets in the reaction cavity flow out of the reaction cavity to the micro-droplet generation structure, and external pressure drives detection separation oil to enter the micro-droplet generation structure from the oil liquid connector; the micro-droplets flowing out of the reaction cavity into the micro-droplet generation structure are separated into queues by the detection separation oil, and the queues enter the fluorescence detection area. According to the invention, through time division multiplexing of a micro-droplet generation structure, droplet generation, amplification and detection are integrated in one chip, and a fully-integrated and fully-closed digital PCR process is realized.
Description
technical field [0001] The invention belongs to the technical field of digital PCR analyzers, and in particular relates to a digital PCR method for an integrated micro-droplet chip. Background technique [0002] Droplet-based microfluidics is a technology platform developed on microfluidic chips in recent years to manipulate small-volume liquids. Its principle is: two immiscible liquids, exemplary , one of which is the oil phase and the other is the water phase. After the oil phase and the water phase enter the microchannel at the same time, under the action of the microchannel, the water phase is distributed in the oil phase in the form of small volume units, forming a A series of discrete micro-droplets. Each droplet acts as a microreactor to complete a set of chemical or biological reactions. [0003] Digital PCR technology is known as the third-generation PCR technology, which has the advantages of absolute quantification and single-molecule detection sensitivity, and ...
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Login to View More IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2563/159C12Q2565/629Y02A50/30
Inventor 苏世圣刘晓彬徐波刘金伟韩应敏周海余杜晓纯杨文军王勇斗
Owner TARGETINGONE TECH (BEIJING) CORP