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Multi-digital PCR chip with high dynamic range and preparation method thereof

A high dynamic range and multiple technology, applied in the field of molecular biology, can solve the problems of limited dynamic range of digital PCR chip detection and insufficient ability of digital PCR chip multiple detection, and achieve the effect of small size, simple operation and simple preparation

Pending Publication Date: 2021-11-19
ZHEJIANG UNIV CITY COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. The digital PCR chip is not capable of multiple detection, 2. The detection dynamic range of the digital PCR chip is limited

Method used

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  • Multi-digital PCR chip with high dynamic range and preparation method thereof
  • Multi-digital PCR chip with high dynamic range and preparation method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0059] The embodiment of the present invention also provides a method for preparing a high dynamic range multiple digital PCR chip, through the following steps:

[0060] (1) Prepare a clean 4-inch silicon wafer;

[0061] (2) adopting standard soft photolithography to make cavity layer 200 molds;

[0062] (3) The mold is spin-coated with a release agent, ventilated and dried;

[0063] (4) Prepare PDMS with A:B=5:1 and pour it on the cavity layer 200 mold, heat and solidify to form the cavity layer 200;

[0064] (5) Prepare PDMS with A:B=10:1 and pour it on the chamber layer 200, and form the support layer 300 after curing;

[0065] (6) The PDMS layer chip is demolded and punched with a puncher to form the inlet and outlet sample ports 228;

[0066] (7) Sealing the chip and the substrate after plasma treatment;

[0067] (8) Paste a layer of polypropylene film on the support layer 300 as the sealing layer 400 to seal the sample inlet and outlet 228 .

Embodiment

[0070] The multiple digital PCR chip with high dynamic range provided by the present invention is prepared through the following steps:

[0071] (1) Silicon wafer preparation: put the clean silicon wafer on a hot plate and bake at 200°C for 4-5 minutes.

[0072] (2) Mold making: Pour a coin-sized SU-8 3025 negative photoresist in the center of the silicon wafer, set the coating program of the homogenizer as follows: 500rpm, 10s; 1200rpm, 30s, the thickness of the photoresist by spin coating is 50 μm. Pre-baking: Bake at 95°C for 15 minutes, then slowly lower to room temperature. Exposure: Align the channel mask with the photoresist-coated silicon wafer, and use a single-sided photolithography machine for exposure, and the exposure time is 6s. Post-baking: bake at 65°C for 1min, bake at 95°C for 5min, and slowly cool down to room temperature. Coating photoresist for the second time: Pour a coin-sized SU-83025 negative photoresist in the center of the silicon wafer, set the c...

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Abstract

The invention belongs to the technical field of nucleic acid detection, and discloses a multi-digital PCR chip with a high dynamic range and a preparation method thereof. The chip comprises a substrate layer, a chamber layer, a supporting layer and a sealing layer which are sequentially bonded from bottom to top, wherein the chamber layer comprises a first-stage sample inlet, multiple detection areas, a second-stage sample inlet, a main channel, a reaction unit and a sample outlet; each detection area comprises a serpentine channel, one end of the serpentine channel is communicated with the first-stage sample inlet; one end of the second-stage sample inlet is communicated with the other end of the serpentine channel; one end of the main channel is communicated with the other end of the second-stage sample inlet; the reaction unit is at least provided with four chambers with different volumes, and the chambers are all connected into the main channel; and the sample outlet is communicated with the other end of the main channel. The multi-detection capability and the dynamic detection range of the digital PCR chip are improved, the dependence on expensive instruments is avoided, and the chip is very suitable for nucleic acid detection scenes.

Description

technical field [0001] The application relates to the field of molecular biology, and relates to a high dynamic range multiple digital PCR chip and a preparation method thereof. Background technique [0002] Microfluidics, characterized by microfabricated structures, typically manipulates fluids on the micron scale, with high throughput and sensitivity. The key elements of microfluidic devices are composed of integrated microchannels, valves, and chambers to achieve efficient fluidic precise control and automatic analysis of complex fluids. As a powerful biomedical device, miniature microfluidic platforms have been applied to tissues engineering, microparticle manufacturing, drug delivery, point-of-care testing (POCT), and tumor screening. With the great advances in microfabrication methods, microfluidic platforms are increasingly used for pathogen detection due to their significant advantages such as low cost, well-controlled microstructure, reduced sample consumption, fas...

Claims

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Application Information

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IPC IPC(8): B01L3/00B01L7/00C12Q1/686
CPCB01L3/5027B01L3/502707B01L7/52C12Q1/686B01L2200/10B01L2300/0887C12Q2537/143C12Q2563/159
Inventor 尹居鑫庄建建夏丽萍
Owner ZHEJIANG UNIV CITY COLLEGE
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