Single molecule DNA fluorescence signal detection system and detection method for array micro-pores

Abstract

The invention provides a single molecule DNA fluorescence signal detection system. The system includes an array chip and an optical detection structure; the array chip is arrayed with multiple array micro-pores and integrated with multiple light-emitting members; and the optical detection structure collects fluorescence signals and converts the fluorescence signals into digital signals so as to realize the detection of single molecule DNAs. A detection method for the array micro-pores is also related. Through the integration of the light-emitting members in the microporous array, the mode of zero-mode waveguide lighting can be avoided, the utilization rates of excitation light can be increased, fluorescence excitation efficiency can be enhanced, and the fluorescence signals can be strengthened; compared with existing blind hole structures of zero-mode waveguides whose bottoms are transparent materials, the loss of light signals passing optical elements can be reduced, and the accuracyrates of fluorescence signal detection and identification can be enhanced; and the size limit of zero-mode waveguides wells can be avoided, and sequencing microporous array chips with higher throughput can be applied, so that single molecule fluorescence sequencing can be realized.

Description

technical field [0001] The invention relates to the field of single-molecule DNA sequencing, in particular to a single-molecule DNA fluorescent signal detection system. Background technique [0002] Single-molecule DNA sequencing technology is a next-generation sequencing technology developed in the past decade, also known as third-generation sequencing technology. It is represented by the SMRT technology of Pacific Bio Corporation in the United States. Zero-mode waveguides wells (ZMWs), a single-molecule DNA polymerase is immobilized in this hole, and the optical signal is introduced into the hole through a zero-mode waveguide. When a certain fluorescently labeled deoxygenated nucleus When a nucleotide is incorporated into a DNA strand, the fluorescence of this specific color persists for a short period of time until a new chemical bond is formed and the fluorophore is cleaved by DNA polymerase, thus passing through four different fluorescently labeled deoxynucleosides. Ac...

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Problems solved by technology

[0005] But at the same time, compared with NGS technology, the third-generation gene sequencing technology based on nanopore chips urgently needs to further improve the detection accuracy of sequencing technology. Low, so the cost of sequencing is high; these factors limit its wide application and development in the field of gene sequencing

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