Digital PCR (polymerase chain reaction) chip and methods for preparing and applying same

A chip and digital technology, applied in the field of digital PCR chip and its preparation, can solve the problems of slow sample injection, low-speed volatilization of water, and difficulty in balancing ventilation requirements, etc., and achieve the effects of fast speed, easy batch manufacturing, and convenient chip punching

Inactive Publication Date: 2018-08-07
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a digital PCR chip and its preparation method and use method, thereby solving the problem that the digital PCR chip in the prior art is difficult to

Method used

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  • Digital PCR (polymerase chain reaction) chip and methods for preparing and applying same
  • Digital PCR (polymerase chain reaction) chip and methods for preparing and applying same

Examples

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Example Embodiment

[0030] Example

[0031] EGFR exon quantification in purified human plasma DNA

[0032] Fabrication of digital PCR chip

[0033] (1) Cleaning of silicon wafers: The silicon wafers were cleaned with Piranha solution, rinsed with deionized water, dried with nitrogen, and baked on a hot plate at 180°C for 30 minutes.

[0034] (2) Fabrication of cavity layer SU8 mold: The silicon wafer was baked on a 180°C hot plate for 30 minutes, then plasma treated for 1 minute, and then spin-coated with SU8 3050 (100μm), followed by photolithography and development to produce an array of supporting microarrays. The pillars serve as a mold for the cavity layer.

[0035] (3) Production of SU8 mold for reaction layer: the silicon wafer is baked on a 180°C hot plate for 30 minutes, then plasma treated for 1 minute, and then spin-coated with SU8 3005 (10 μm), photolithography and development are performed to produce a reaction chamber layer reaction The communication branch between the cavity and...

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Abstract

The invention provides a digital PCR (polymerase chain reaction) chip and methods for preparing and applying the same. The digital PCR chip comprises a glass support layer, a seal layer, a reaction layer, a cavity layer and a waterproof layer. The glass support layer, the seal layer, the reaction layer, the cavity layer and the waterproof layer are sequentially arranged from bottom to top, the reaction layer and the seal layer are bonded to form main flow channels and a plurality of independent PCR chambers between the reaction layer and the seal layer, the PCR chambers are separated from oneanother by the main flow channels, the cavity layer and the reaction layer are bonded to form cavities between the cavity layer and the reaction layer, sample inlets and sample outlets which are sequentially perforated are further formed in the reaction layer, the cavity layer and the waterproof layer, and cavity layer inlets and cavity layer outlets which are sequentially perforated are formed inthe cavity layer and the waterproof layer; the seal layer, the reaction layer and the cavity layer are made from polydimethylsiloxane, and the waterproof layer is made from parylene. The digital PCRchip and the methods have the advantage that the digital PCR chip is easy to operate, high in speed, flux and automation degree and low in manufacturing cost.

Description

technical field [0001] The invention relates to the field of molecular biology, and more particularly to a digital PCR chip and a preparation method and a use method thereof. Background technique [0002] Polymerase chain reaction (Polymerase Chain Reaction, PCR) is a molecular biology technique for amplifying and amplifying specific DNA fragments, which can be regarded as special DNA replication in vitro. It has been widely used in the fields of molecular biology such as genetic testing, gene amplification, and genetic engineering, and has played an irreplaceable role in clinical medicine, forensic science, paternity testing, and environmental testing. However, the PCR reaction is exponentially amplified, and can be amplified millions of times within a few tens of minutes, and it is difficult to determine the content of the original PCR template from the PCR product. In order to accurately quantitatively analyze the nucleic acid content, digital PCR (digital PCR, dPCR) tec...

Claims

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Application Information

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IPC IPC(8): C12M1/38C12M1/00
CPCB01L3/502784B01L7/52B01L2300/0861C12Q1/6851C12Q2531/113
Inventor 徐铁刚吴蕾王雪凤李昕欣
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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