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Nucleic acid sequencing device and system

A nucleic acid sequencing and detection device technology, applied in the field of pyrosequencing-based nucleic acid sequencing devices and nucleic acid sequencing devices, can solve the problem of separation and detection results accuracy interference, inflexible DNA single-strand collection methods, and high cost of DNA single-strand separation. problems, to avoid a single product structure, enrich the structure and variety, and ensure the quality and efficiency.

Inactive Publication Date: 2017-05-31
武汉菲思特生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

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  • Nucleic acid sequencing device and system
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  • Nucleic acid sequencing device and system

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0096] Figure 1 to Figure 3 The first embodiment of the nucleic acid sequencing device of the present invention is shown. The nucleic acid sequencing device of the present invention can be used for pyrosequencing detection and analysis of DNA sequences, the DNA sequence to be sequenced is a target sequence, and pyrosequencing is performed after the target DNA sequence is amplified.

[0097] The nucleic acid sequencing device of this embodiment includes a sample area, a reaction area, a detection area and a control area. The sample area, reaction area and detection area are installed on the analysis table 6, and the sample area, reaction area and detection area are monitored and controlled by the control area. The whole analysis process is operated by manipulator.

[0098] The sample area includes an electrophoresis apparatus 1 and a separation disc 2 . The electrophoresis instrument 1 is provided with a gel platform and light-emitting diodes, and agarose beads are arranged...

Embodiment 2

[0149] Because a chain collected on the filter membrane 22 needs to be sucked out or poured out, such a collection method may cause a certain collection loss, so the inventor preferably arranges the filter column 21 as a double-head structure with two ends exchanged, and the filter membrane 22 is located on the non-end surface of the filter column 21. When collection is required, replace the two ends of the filter column 21 and use conventional methods, such as centrifugation, to elute all DNA single strands on the filter membrane 22 to reduce sample loss.

[0150] Such as Figure 5 As shown, the difference between this embodiment and embodiment 1 is that the filter column 21 in embodiment 2 is a hollow cylinder with a symmetrical structure up and down, and the filter membrane 22 is vertically located in the middle of the hollow cylinder, and the filter column 21 can be For replacement use, the inner diameter of the hollow cylinder is 4.5mm, and the diameter of the filter memb...

Embodiment 3

[0154] Such as Figure 6 As shown, the difference between this embodiment and Example 1 is that the filter column 21 of the DNA single-strand separation device in Example 3 adopts a space truncated circular structure, and the filter membrane 22 is arranged on the common top surface of the two circular truncated filter columns 21 , the diameter of the top surface of the circular truncated filter column 21 is consistent with the diameter of the filter membrane 22, and the two sides of the filter membrane 22 are provided with a pressing mechanism (not shown in the figure), so that the fixed filter membrane 22 cannot be displaced. The side wall of the opening lower end of the frustum-shaped filter column is provided with a boss to ensure that it can be fixed on the boss of the collecting pipe 23 when the two ends are reversed.

[0155] The separation method using the separation device belonging to this embodiment is the same as the step described in embodiment 1, and the differenc...

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Abstract

The invention provides a nucleic acid sequencing method and a device. The device comprises a sample area, a reaction area, a detection area and a control area, wherein the sample area comprises an electrophoresis apparatus and a rotatable separating plate; the separating plate comprises at least one DNA separating area; the separating area comprises hollow filtering columns in which a filtering film is arranged; a DNA single chain which is connected with an affine connector and a 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 nucleic acid sequencing device provided by the invention has the advantages of simple and convenient operation and quick detection.

Description

technical field [0001] The invention relates to a nucleic acid sequencing device, in particular to a pyrosequencing-based nucleic acid sequencing device, which belongs 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 enzymes. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34
CPCC12N15/1017C12Q1/6806C12Q1/6827C12Q1/6869C12Q2565/301C12Q2535/122C12Q2523/32C12Q2565/631C12Q2563/107
Inventor 陈立波刘丹孙悦
Owner 武汉菲思特生物科技有限公司
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