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

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...

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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 achieve a balance between the necessary ventilation requirements and the airtight requirements to prevent moisture evaporation, Causes the problem of slow sample injection and low-speed evaporation of water

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