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A high-density array chip device and application for digital nucleic acid amplification

A nucleic acid and chip technology, applied in the field of detection devices in many fields, can solve the problems of unfavorable preparation methods and application costs, difficulty in entering ordinary laboratories, expensive chips and supporting instruments, etc., to reduce experimental costs, prevent external pollution, The effect of preventing cross-contamination

Active Publication Date: 2011-12-14
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fabrication of this digital PCR chip based on PDMS elastic microvalve involves the alignment and sealing of multilayer structures, and the fabrication process is long and complicated.
Since the detection device used is a high-precision digital CCD, the entire commercialized chip and supporting instruments are very expensive, and the operation is complicated, and some require special instruments, which are expensive and difficult to enter in ordinary laboratories health screening method to promote
Therefore, from the perspective of large-scale application prospects of digital PCR chip technology, the current preparation method and application cost are unfavorable.

Method used

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  • A high-density array chip device and application for digital nucleic acid amplification
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  • A high-density array chip device and application for digital nucleic acid amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 High-density glass chip device for digital nucleic acid amplification

[0035] Referring to Figures 1-3, an integrated flow chip device for digital nucleic acid amplification is composed of a vacuum system 1, a pipeline 2, a suction cup 3, a capping layer 7 and a reaction layer 10, and the capping layer 7 and the reaction layer After the layers 10 are sealed, an integrated flow path chip assembly is formed. The two ends of the capping layer 7 are provided with a sample inlet 9 and a sample outlet 5. The reverse side of the capping layer 7 is an irregular hexagonal concave surface. When the capping layer After 7 is sealed with the reaction layer 10, the channel 6 is formed. The reaction layer 10 is provided with high-density integrated reaction chambers 11 arranged in a matrix. The vacuum system 1 is connected to the suction cup 3 through the pipeline 2, and the suction cup 3 is placed on the cover layer 7 for sampling. On the port 5, the sample outlet 5 is con...

Embodiment 2

[0041] Example 2 High-density monocrystalline silicon chip device for digital nucleic acid amplification

[0042] The structure of the device is the same as in Example 1.

[0043] The reaction layer 10 is made of a polished single-crystal silicon wafer with a thickness of 500 μm, and the reaction chamber 11 is formed by photolithography and chemical wet etching. The reaction chamber 11 is a square with a side length of 100 μm, 200 μm or 300 μm, and a depth of 50 μm, 100 μm, 200 μm, 300 μm. According to different widths and etching depths, the volume of the reaction chamber 11 can be 500 pL, 1 nL, 2 nL, 4.5 nL, 8 nL, 9 nL, 18 nL; the reaction chamber 11 can also be pentagonal, rhombus, circular shape or other polygons.

[0044] The cover layer 7 can be made of glass or a transparent PMMA plate according to different applications. Use standard photolithography and etching techniques to form an irregular hexagonal concave surface on the reverse side of the capping layer 7, wit...

Embodiment 3

[0049] Example 3 Digital-Loop Mediated Isothermal Amplification (Digital-Loop Mediated Isothermal Amplification, Digital-LAMP)

[0050] The device of Example 1 is used to inject samples, and then perform subsequent digital nucleic acid loop-mediated isothermal amplification.

[0051] Specific steps:

[0052] 1. The channel structure is the same as that in Embodiment 1, and the designed structure is printed onto film with an inkjet printer as a mask for photolithography.

[0053] 2. The reaction layer 10 is made of glass with a thickness of 1 mm, and the reaction chamber 11 is made by standard photolithography and etching technology. The reaction chamber 11 is a square with a side length of 100 μm, 200 μm or 300 μm, and a depth of 50 μm or 100 μm. or 200 μm; according to different widths and etching depths, the volume of the reaction chamber 11 can be 500 pL, 1 nL, 2 nL, 4.5 nL, 8 nL, 9 nL or 18nL; the reaction chamber 11 can also be pentagonal, Rhombus or circle.

[0054] 3...

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Abstract

The invention provides an integrated flow path chip device used for digital nucleic acid amplification. The device is composed of a vacuum system, a pipeline, a sucker, a sealing cover layer and a reaction layer, wherein two ends of the sealing cover layer are respectively provided with a sample inlet and a sample outlet; a path is formed between the sealing cover layer and the reaction layer; the reaction layer is provided with small reaction chambers; the sucker is arranged on the sample outlet; the vacuum system is connected with the sucker via a pipeline; and the sample inlet and the sample outlet are sealed by adhesive tape paper. According to the integrated flow path chip device, trace reaction samples can be distributed to thousands of small reaction chambers, and mineral oil is led in via a path in the sealing cover layer to separate thousands of small reaction chambers so as to realize the single molecule amplification and precisely quantify the nucleic acid molecules. The device is reasonable to design and is a minitype device used for digital nucleic acid amplification, which has low cost and is easy to operate.

Description

technical field [0001] The invention belongs to detection devices in multiple fields such as life sciences and medicine, and relates to an integrated flow path chip device for digital nucleic acid amplification and its application. Background technique [0002] With the rapid development of science and technology, especially the development of life sciences, the research and application of biomedicine have gone from the overall and cellular level to the molecular level, and the research on the diagnosis and treatment of many diseases has gradually entered the gene level. As one of the most important biomacromolecules in the living body, nucleic acid is the most critical component in living cells. It carries genetic information and is the material basis of inheritance. Although the final product of gene expression is protein, it is nucleic acid that determines the structure of proteins and enzymes. Therefore, the systematic and effective detection and analysis of nucleic aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12Q1/68
Inventor 牟颖朱强远高一博于丙文金伟
Owner ZHEJIANG UNIV
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