DNA sequencing device and system based on pyrosequencing

Abstract

The invention provides a DNA sequencing system based on pyrosequencing. The DNA sequencing system comprises a sample area, a reaction area, a detection area and a control area, wherein the sample area comprises a rotatable separating plate; the separating plate comprises at least one DNA separating area; the separating area comprises a hollow filtering column in which a filtering film is arranged; a DNA single chain which is connected with an affine connector and the DNA single chain which is not connected with the affine connector are filtered by the filtering film and then are separated; the reaction area comprises a sampling part and a sample slot; the sampling part comprises a dNTP reagent slot, a sampling needle frame and a plurality of sampling needles arranged on the sampling needle frame; the sampling needle frame is located on the sample slot; a plurality of slot positions are formed on the sample slot; the detection area comprises a bioluminescence detection device; the slot positions are connected with the bioluminescence detection device; the sample area, the reaction area and the detection area are monitored and controlled through the control area. The DNA sequencing system and device based on pyrosequencing, provided by the invention, have the advantages of simple and convenient operation and quick detection.

Description

technical field [0001] The invention relates to a pyrosequencing-based DNA sequencing device, in particular to a pyrosequencing-based DNA sequencing device and system, belonging to the field of gene detection. Background technique [0002] 1. Overview of Pyrosequencing [0003] Pyrosequencing is a sequencing technology developed in 1987 based on the detection of pyrophosphate (PPi) released during DNA synthesis. The pyrosequencing reaction is catalyzed by a series of enzymes. Visible light proportional to the polymerization number of deoxynucleoside triphosphate (dNTP) is generated, and the purpose of determining the DNA sequence can be achieved by detecting visible light. There are two implementation methods of pyrosequencing: liquid phase pyrosequencing (Liquid Phase Pyrosequencing) and solid phase pyrosequencing (Solid Phase Pyrosequencing). [0004] Liquid-phase pyrosequencing is an enzyme cascade chemiluminescent reaction in the same reaction system catalyzed by four ...

AI Technical Summary

Problems solved by technology

Generally, 24 (4*6) suction filter needles are used as a group. When using it, it is necessary to ensure that there are sufficient samples or reagents to ensure the normal operation of the workstation. Therefore, this method of collecting DNA single strands is very inflexible and can only be used A fixed amount is added to the workstation to work, and a large amount of loss occurs in the process of multiple suction filtration and transfer, which is very unfavorable for micro collection, and the suction filter needle group needs to work at the same time, which has a certain volume for each component of the workstation requirements, the entire workstation takes up a lot of space

The huge system makes the micro-separation column need to pipette back and forth repeatedly during the DNA single-strand separation operation. The operation is very cumbersome, not only the separation cycle is long, the efficiency is low, and the overall equipment price is relatively expensive, resulting in the high cost of DNA single-strand separation. It also needs to consume a lot of reagents and other resources, which is extremely uneconomical

In addition, the suction filter needle in this workstation is made of metal, which is expensive and is often reused after treatment, so it is easy to cause cross-contamination between residues, the reliability is not high, and it will affect the accuracy of separation and detection results. Certain interference and influence

And during the solution extraction process, some residual solution will stick to the wall, so that a certain amount of target DNA single strands cannot be adsorbed by the micro-separation column, resulting in a decrease in the proportion of obtained DNA single strands, affecting the separation rate and causing waste

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