Sample adding device for pyrosequencing

Abstract

The invention provides a sample adding device for pyrosequencing. The sample adding device comprises a sample area and a reaction area; the sample area comprises a rotational separating disc, the separating disc comprises at least one DNA separating area, the separating area comprises a hollow filtering column with the inside provided with a filtering film, and DNA single chains connected with compatible connecting bodies and DNA single chains unconnected with compatible connecting bodies are filtered through the filtering film to be separated; the reaction area comprises a sample adding part and a sample trough, and the sample adding part comprises a dNTP reagent trough, a sample adding needle frame and a plurality of sample adding needles installed on the sample adding part; the sample adding needle frame is located on the sample trough; a plurality of trough positions are arranged on the sample trough. The sample adding device and system for pyrosequencing have the advantages of being easy and convenient to operate and rapid in detection.

Description

technical field [0001] The invention relates to a sampling device for pyrosequencing, in particular to a pyrosequencing-based sampling device and system for pyrosequencing, 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 ...

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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