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Digital PCR chip based on seaweed gel liquid drop and application thereof

A technology of algae glue and droplets, applied in the field of microfluidics, can solve the problems of reducing integration, increasing operation time, increasing instrument cost, etc., and achieves the effects of good stability, improved detection efficiency, and detection sensitivity.

Active Publication Date: 2017-05-31
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bio-Rad's droplet PCR instrument operates differently from droplet generation, nucleic acid amplification, and fluorescence detection, which increases the complexity of the operation, and each step brings the loss caused by sample transfer, reducing the Integration, which increases both operating time and instrument cost

Method used

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  • Digital PCR chip based on seaweed gel liquid drop and application thereof
  • Digital PCR chip based on seaweed gel liquid drop and application thereof
  • Digital PCR chip based on seaweed gel liquid drop and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Droplet capture chip

[0088] Such as figure 1 As shown, a droplet capture chip 1 includes a plurality of fluid microchannels 11 arranged in parallel, wherein a plurality of droplet capture structures 12 are formed at intervals along the fluid microchannel 11, the droplet capture structures 12 includes a droplet entry end 121 and a droplet suppression end 122. The suppression droplet outflow end 122 is connected to the adjacent fluid microchannel; for ease of control, all droplet capturing structures 12 have the same size and orientation. The two ends of each fluid microchannel 11 are respectively provided with a first inlet 13 and a first outlet 14; droplets enter the fluid microchannel 11 from the first inlet 13 and move from left to right under pressure; A plurality of second inlets 15 and second outlets 16 arranged in parallel are respectively provided on both sides of the fluid microchannel. The second inlets 15 and second outlets 16 are used to apply pressure perpen...

Embodiment 2

[0099] Nucleic acid amplification reaction method

[0100] A nucleic acid amplification reaction method, including:

[0101] (1) Provide the digital PCR chip based on alginate droplets in Example 2;

[0102] (2) Add a dispersed phase and a continuous phase to the first liquid inlet and the second liquid inlet of the digital PCR chip respectively, and the dispersed phase contains algin monomer (2wt%), PCR reaction solution, calcium carbonate nano Granules (40mmol / L), KCl (7mmol / L) and NaCl (75mmol / L), the continuous phase is mineral oil containing 2μL / mL acetic acid and 2.5% span 80 surfactant;

[0103] The dispersed phase and the continuous phase respectively flow through the first and second inlet channels of the digital PCR chip. After contact, droplets of alginate are generated in the droplet generation area, and then the droplets of alginate are captured into the liquid through the fluid microchannel. Droplet capture chamber;

[0104] (3) A temperature control module is placed u...

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Abstract

The invention discloses a liquid drop capturing chip. The liquid drop capturing chip comprises a plurality of fluid micro channels arranged in parallel, wherein a plurality of liquid drop capturing structures are formed along the fluid micro channels at intervals; the liquid drop capturing structures comprise liquid drop entering ends and inhibition liquid drop flowing-out ends; and the inhibition liquid drop flowing-out ends are communicated with the adjacent fluid micro channels. The invention further discloses a fully integrated digital PCR chip based on a seaweed gel liquid drop and application thereof. The digital PCR chip based on the seaweed gel liquid drop, disclosed by the invention, can finish the full process from the generation of liquid drops to capture of the liquid drops and to the detection of the liquid drops. And moreover, seaweed gels are introduced; after the seaweed gels are solidified, the seaweed gel liquid drops are stable, cannot be fused and cannot affect the PCR amplification efficiency; and the method can be applied to the detection of circulating tumor DNA in the blood and other related biology and medical sciences; and due to the introduction of the seaweed gels, the application can be further expanded to single cell analysis and so on.

Description

Technical field [0001] The invention belongs to the technical field of microfluidics, and particularly relates to a digital PCR chip based on seaweed gel droplets and its application. Background technique [0002] Microfluidic technology is one of the cutting-edge fields of science and technology with multiple disciplines. It has a series of characteristics such as small size, high efficiency, high integration, fast analysis speed, and low price. Droplet microfluidics is an important branch of microfluidic technology. The microfluidic droplet system provides a potential tool for single cell analysis. Using the microfluidic system, single cells can be quickly segmented and high-throughput parallel observations Detection. The use of microfluidic droplets for cell culture is already a relatively mature technology. The use of specific microchannel designs such as capture arrays, culture chambers, and microwell arrays to fix microdroplets can achieve online cell culture. The relativ...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12Q1/68
CPCB01L3/502784B01L7/52B01L2300/0861C12Q1/686C12Q2563/159C12Q2543/101
Inventor 吴天准李丽军於林芬
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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