Nanopore sequencer based on nano nuclease and sequencing method thereof

Abstract

The invention relates to the technical field of gene sequencing, in particular to a nanopore sequencer structure based on nano nuclease and a sequencing method thereof. The sequencer comprises a power source, a liquid pool and a substrate placed in the liquid pool, a groove is formed in the substrate, a through hole penetrating through the substrate is drilled in the bottom of the groove, the area of the bottom of the groove is larger than the aperture of the through hole, an opening of the groove is covered with a nanopore chip, nanopores of the nanopore chip are located in a notch of the groove, an electromagnet is arranged on the side, opposite to the groove, of the substrate, the electromagnet is close to the through hole, nano nucleic acid hydrolase is placed in the groove, the nano nucleic acid hydrolase is attracted by the electromagnet to be fixed at the bottom of the groove, the negative electrode of the power source is inserted into or close to the through hole, and the positive electrode of the power source is close to the nanopores. The nanopore sequencer has the advantages that the sequencing precision is high, operation is convenient, the sequencing method can achieve sequencing by opening the power source, other operations of capturing a DNA single chain to be detected are not needed, operation is easy, and efficiency is high.

Description

technical field [0001] The invention relates to the technical field of gene sequencing, in particular to a nanopore sequencer based on nanonuclease and a sequencing method thereof. Background technique [0002] At present, solid-state nanopore sequencing technology has been widely used in gene sequencing. The solid-state nanopore sequencing technology allows the DNA strand to be tested to pass through the nanopore of the nanopore chip, and the nanopore collects the base signal on the DNA to be tested to achieve DNA sequencing. [0003] At the same time, there are many challenges in solid-state nanopore sequencing technology. Mainly include: 1. Time resolution. Under the action of electric field, the speed of DNA passing through the hole is too fast to collect enough effective data points; 2. Spatial resolution. The distance between adjacent bases is only 0.34nm. To achieve single base The thickness of the resolution nanopore must be less than 0.34nm, and it is necessary to ...

AI Technical Summary

Problems solved by technology

However, there are certain problems in these methods: the optical tweezers system needs to rely on a complex optical system, and can only control a single DNA molecule at a time, and cannot achieve parallel detection; it is difficult to avoid the serious interference of the tuning fork vibration to the detection signal by using magnetic tweezers

If you increase the ion concentration, the noise will increase significantly or the amplitude of the detection signal will decrease; if you add glycerol to the solution to increase the concentration of the solution, while slowing down the movement of DNA molecules, it will also slow down the movement of ions, which will also make The signal to block the current becomes smaller; in the method of modifying the nanopore, the strong interaction of DNA with the nanopore leads to the blockage of the nanopore

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