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Integral rigid runner droplet digital PCR system and method

An integrated, rigid technology, applied in chemical instruments and methods, biochemical equipment and methods, biochemical instruments, etc., to achieve the effects of reducing sample contamination, droplet fusion, and improving thermal conductivity

Inactive Publication Date: 2017-03-22
UNIV OF SCI & TECH OF CHINA
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] The technical problem of the present invention is to overcome the deficiencies of the prior art, provide an integrated rigid flow channel droplet digital PCR system and method, have a highly integrated design, integrate a droplet generating device (2) with strong pressure bearing capacity and adopt rigid Thermal circulation flow channel device, with high thermal conductivity, can effectively inhibit droplet breakage and cross-contamination between samples

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  • Integral rigid runner droplet digital PCR system and method
  • Integral rigid runner droplet digital PCR system and method
  • Integral rigid runner droplet digital PCR system and method

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[0022] Such as figure 1 As shown, the present invention includes a power device 1 , a droplet generating device 2 , a droplet PCR reaction module 3 , a fluorescence detection controller 4 and a PC control module 5 . In this example, mineral oil added with EM90 was used as the continuous phase fluid, and the sample containing the target DNA template was used as the dispersed phase fluid to generate monodisperse droplets containing template molecules. Among them, the volume fraction of EM90 contained in mineral oil is 2%, and the specific proportion of the sample containing DNA template is as follows: Each 1mL PCR Mix reaction reagent contains 873 μL ddH2O, 100 μL 10*Buffer, 20 μL dNTP Mix, 0.9 μL Lib5S1, 0.9 μL Lib3A2 and 5μL Pfu enzyme 5U / mL, after the above ratio is completed, add 4% EvaGreen dye to it. After the dye can bind to a specific template, it will produce fluorescence after being excited by light of a specific wavelength. The fluorescence intensity can be used To ...

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Abstract

The invention relates to an integral rigid runner droplet digital PCR system. The system comprises a power device, a droplet generation device, a rigid thermal circulation runner, a thermal circulation temperature control module, a fluorescent detection controller and a PC control module. A rigid thermal circulation pipeline has high heat conducting performance, and a rigid wall surface has low roughness and small relative deformation. The integral rigid runner droplet digital PCR system can be used for online detecting a droplet in real time by adopting the droplet generation device with a high loading capacity and a terminal fluorescent detection system, and has the characteristics of flexible temperature control, safety, no need of a special PCR reaction chip, high fluorescent detection flux, high sensitivity and the like; and the system can avoid the problems of measurement accuracy reduction and the like caused by droplet breakage and mutual fusion among droplets since the traditional droplet digital PCR equipment needs to manually transfer droplets in different reaction modules. The whole system forms a highly-integrated design by droplet generation, PCR reaction thermal circulation and terminal droplet fluorescent real-time online detection.

Description

technical field [0001] The invention relates to an integrated rigid channel droplet digital PCR system and method, which belongs to the field of medical equipment and is suitable for polymerase chain reaction (PCR) requiring high throughput, on-line detection and complete quantification of amplified templates. Background technique [0002] Polymerase chain reaction (PCR) is a very important method for nucleic acid analysis. Thanks to its ability to quickly enrich target molecules and prepare for the next step of analysis, it is widely used in the fields of medical and biological research, greatly accelerating The research process of biological genome structure. Due to the large volume of liquid, traditional PCR instruments require complex heating and cooling cycles, consume a lot of energy, lack real-time monitoring and cannot perform absolute quantitative analysis. Digital PCR technology is a new type of PCR technology that has developed rapidly in recent years, and it can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12Q1/68
CPCB01L7/52B01L2300/1805C12Q1/686
Inventor 陈进宇何立群罗昭锋
Owner UNIV OF SCI & TECH OF CHINA
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