High-integration equidistance equipartition nucleic acid amplification micro-fluidic chip and application

Abstract

The invention provides a high-integration equidistance equipartition nucleic acid amplification micro-fluidic chip. The high-integration equidistance equipartition nucleic acid amplification micro-fluidic chip sequentially consists of a cover glass layer, a reaction layer, a decorative layer and a sealing layer from bottom to top in a thermal bonding manner, wherein the reaction layer is provided with n reaction modules, and each reaction module is provided with a circulation passage, a reaction micro-cavity and a sample feeding opening by utilizing a multilayer soft photoetching technology. By adopting the chip, the circulation distance of the sample before being introduced into a small chamber is identical, the sample feeding quantity is identical, and an error caused by the non-uniformity of the reaction volume can be avoided. The experiment cost is reduced. The sample feeding speed is increased. Quantitative detection in real time can be realized. The chip is reasonable in design, simple in structure, simple to operate and low in production cost and can be applied in the real-time fluorescent quantitative nucleic acid amplification, digital nucleic acid amplification, single-cell separation of nucleic acid amplification, SNP detection, detection of single-base mutation, detection of copy number imbalance, research of single-cell gene expression profile, early detection of cancer marker, stem cell differentiation identification, cell separation identification and the like.

Description

technical field [0001] The invention belongs to a detection device in the field of biology, and relates to a highly integrated microfluidic chip for nucleic acid amplification for equally spaced and equally divided samples, in particular to a highly integrated equidistant and equally divided nucleic acid amplification microfluidic chip, and Its application of this chip. Background technique [0002] Major diseases such as cancer, cardiovascular and cerebrovascular diseases, and infectious diseases claim the lives of millions of people in my country every year. The individual differences between disease diagnoses have increased the demand for high-quality and efficient personalized medical care. New prevention and diagnosis methods Method research is becoming more and more urgent. The occurrence of diseases is generally caused by molecular mutations, and it is of great importance and urgency to realize early diagnosis at the molecular level. Therefore, it is of great signifi...

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Problems solved by technology

[0006] Many research groups have developed digital PCR platforms, such as microfluidic chips, microemulsion droplets, centrifugal chips, slipchips, etc., but these platforms are complex to operate, and some require special instruments, which are expensive, such as microvalve-controlled microchips. Fluidic chips require a control system; droplet chips require a droplet generator; some can only be injected once, which means that the upstream sample pretreatment of digital PCR, including DNA extraction and transcription, can only be done outside the chip. The results are greatly affected; most microfluidic chips need to design a sample port in addition to the sample port, resulting in sample waste and aerosol pollution; these have brought difficulties to the application of ordinary laboratories, and it is even more difficult to be used as a routine health screening tool

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