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A kind of preparation and modification method of digital pcr chip

A chip and digital technology, applied in the field of digital PCR chip preparation and modification, can solve the problems of high cost, complicated digital PCR chip processing technology, low chip throughput, etc., to reduce processing costs, improve hole efficiency, and reduce processing The effect of the process

Active Publication Date: 2020-07-10
宁波胤瑞生物医学仪器有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention provides a method for preparing and modifying a digital PCR chip in order to solve the problems of complex digital PCR chip processing technology, high cost, low throughput of the chip, and ease of use of the chip in the prior art. The specific technical scheme is as follows:

Method used

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  • A kind of preparation and modification method of digital pcr chip
  • A kind of preparation and modification method of digital pcr chip
  • A kind of preparation and modification method of digital pcr chip

Examples

Experimental program
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Embodiment 1

[0037] A kind of preparation method of digital PCR chip, concrete steps are as follows:

[0038] Step 1: Select silicon material as the base layer of the chip;

[0039] Step 2: After polishing one side of the base layer of the chip, the polymer material is embedded on the polished surface of the base layer of the chip at 65°C using a laminating machine, and then the protective film on the surface of the polymer material is removed to form a polymer material panel; The thickness of the molecular material is 25 μm; the polymer material is a photosensitive thick epoxy resin sheet, and the photosensitive thick epoxy resin sheet is composed of a photoresist containing cation-cured modified epoxy resin; the photoresist of the epoxy resin contains no antimony photoacid generator and is prepared in a strictly controlled solvent-free process;

[0040] Step 3: Baking the polymer material panel in step 2 at 95°C for 10 minutes, and then exposing on the exposure machine to form a close-p...

Embodiment 2

[0047] Compared with Example 1, the thickness of the polymer material in this example is 20 μm, and in the regular hexagonal pattern array, the distance D between the opposite sides of the regular hexagon is 0.03mm, and the gap d of the regular hexagon is 0.013mm; The widths a2 and a3 of the upper and lower grooves are 0.02775 mm, and the widths a4 and a1 of the left and right grooves are 0.0248 mm. The modification method of digital PCR in this embodiment is:

[0048] S1, the chip obtained in step 5 is subjected to plasma treatment by a plasma machine to obtain chip A,

[0049] S2. Soak the chip A in step S1 in bovine serum albumin, then dry it and use it directly.

[0050] The concentration range of bovine serum albumin in the step S2 is 10 mg / mL.

Embodiment 3

[0052] Compared with Example 1, the thickness of the polymer material in this embodiment is 15 μm, and in the regular hexagonal pattern array, the distance D between the opposite sides of the regular hexagon is 0.022mm, and the gap d of the regular hexagon is 0.013mm; The widths a2 and a3 of the upper and lower grooves are 0.02825 mm, and the widths a4 and a1 of the left and right grooves are 0.02015 mm.

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Abstract

The invention provides a preparation of a digital PCR chip and a modification method thereof. The preparation method comprises the following steps: selecting a silicon material as a chip base layer; polishing a single surface of the chip base layer, embedding a high polymer material into the polished surface of the chip base layer, and removing a protective film on the surface of the high polymermaterial to form a high polymer material panel; baking the high polymer material panel in the above step, and exposing on an exposure machine to form a densely arranged pattern array of reaction micropores on the surface; soaking the densely arranged pattern array obtained in the step 3 in a developing solution, and then drying the densely arranged pattern array; and scribing the high polymer material panel dried in the above step to complete the preparation of the digital PCR chip. The modification method comprises the following steps: carrying out plasma treatment on the digital PCR chip through a plasma machine, and then carrying out digital PCR reaction. The problems that in the prior art, a digital PCR chip is complex in machining process, high in cost, low in chip flux, easy to use and the like are solved.

Description

technical field [0001] The invention belongs to the technical field of PCR, and in particular relates to a method for preparing and modifying a digital PCR chip. Background technique [0002] Digital PCR (digital polymerase chain reaction, dPCR) generally includes two parts, namely PCR amplification and fluorescence signal analysis. In the stage of PCR amplification, digital PCR first dilutes the sample to the single-molecule level, and then evenly distributes it to hundreds to tens of thousands of units for reaction. Different from real-time fluorescent quantitative PCR (real time quantitative PCR, qPCR) method of real-time fluorescence measurement for each cycle, digital PCR collects the fluorescence signal of each reaction unit after the amplification is completed, and the fluorescence signal is recorded as 1 , no fluorescent signal is recorded as 0, and the reaction unit with fluorescent signal contains at least one copy. Theoretically, when the target DNA concentratio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851C12M1/34C12M1/00
CPCC12Q1/6851C12Q2531/113C12Q2563/137C12Q2549/125
Inventor 隋硕金鑫浩刘一博任鲁风张未来俞育德于军
Owner 宁波胤瑞生物医学仪器有限责任公司
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