WO2018214907A1

WO2018214907A1 - Device and method for nucleic acid extraction and purification

Info

Publication number: WO2018214907A1
Application number: PCT/CN2018/087999
Authority: WO
Inventors: 彭年才; 李明; 苗保刚; 李政; 田春; 孙瑶
Original assignee: 西安天隆科技有限公司
Priority date: 2017-05-24
Filing date: 2018-05-23
Publication date: 2018-11-29

Abstract

The present invention relates a device and a method for nucleic acid extraction and purification. Essential reagents of nucleic acid extraction are respectively packaged, combined, and integrated in a closed system. Under an external force applied by a supporting apparatus, the nucleic acid extraction and purification and subsequent detection and analysis processes are carried out in sequence. The nucleic acid extraction process is simplified, extraction is sped up, and cross-contamination between samples is avoided at the same time. Also, the complex processing of nucleic acid extraction automatically performed from one step to the next is achieved.

Description

Nucleic acid extraction and purification device and method

Technical field

The invention belongs to the field of nucleic acid detection, and particularly relates to a nucleic acid extraction and purification device and a method, which complete fully automatic sample nucleic acid extraction, amplification and detection in a matched instrument.

Background technique

Nucleic acid is a biomacromolecular compound that is polymerized from many nucleotides. It is a carrier of genetic information of living organisms and is one of the most basic substances in life. Therefore, whether it is the study of nucleic acid sequence and structure, the exploration of gene function and expression, or the genetic engineering or protein engineering, the nucleic acid is first isolated and purified. At the same time, the quality of nucleic acid extraction products will also be directly It is related to the success or failure of subsequent experiments. Therefore, the extraction and purification of nucleic acids is the most important and basic operation in molecular biology experimental techniques.

Nucleic acid extraction and purification methods are diverse and can involve a variety of different processing methods, including biological, chemical, mechanical, thermal or acoustic. However, no matter what method is used for treatment, it basically includes the main steps of cell release, nucleic acid isolation and purification, precipitation or adsorption of nucleic acid, and release of nucleic acid in buffer, and these steps are almost every step. The need for a combination of different reagents, the sample reacts differently between the various reagents, thereby achieving the final separation and purification of the nucleic acid. These steps are numerous and complex, and traditional methods are manually processed by professionals in the laboratory. However, these manual methods are inefficient, costly, and subject to operational errors.

With the emergence of nucleic acid extraction equipment of different degrees of automation, laboratory personnel can use nucleic acid extraction equipment to simultaneously process multiple different samples, but how to package, integrate and integrate the reagents necessary for nucleic acid extraction into a closed system. It simplifies the nucleic acid extraction process, speeds up the extraction, avoids cross-contamination between samples, and realizes complex processing devices and methods that are automatically implemented from one step to the next, which is still expected by the market and urgently needs to be developed. of.

Summary of the invention

In order to solve the problems of low efficiency, high cost, complicated operation and the like in the conventional method for extracting and purifying nucleic acid, the present invention provides a nucleic acid extraction and purification device and method, based on a three-dimensional double-sided disc type sealing device with a certain thickness, and is matched Under the external force applied by the instrument, the nucleic acid extraction and purification and subsequent detection and analysis processes are carried out in sequence. Further, in order to reduce the processing difficulty and shorten the device

Claims

In the processing cycle, the present invention also provides another nucleic acid extraction and purification device.

The technical solution of the present invention is to provide a nucleic acid extraction and purification device, which is characterized in that it comprises a substrate, a plurality of reagent chambers disposed on the substrate, a nucleic acid adsorption chamber, a waste liquid chamber, a product chamber, and In the nucleic acid analysis unit, the reagent chamber, the waste liquid chamber and the product chamber communicate with the nucleic acid adsorption chamber through corresponding flow channels, and the waste liquid chamber and the product chamber communicate with the respective reagent chambers through corresponding return air passages. Each of the chambers, the flow channels, and the return air passages are located on a front surface, a reverse surface, or a through-substrate of the substrate, wherein the reagent chamber is disposed near a center of the substrate; and the reagent chamber, the nucleic acid adsorption chamber, the waste liquid chamber, and the nucleic acid An analysis unit, or the reagent chamber, the nucleic acid adsorption chamber, the product chamber, and the nucleic acid analysis unit are sequentially disposed from the inside to the outside in a radial direction of the substrate, wherein the nucleic acid analysis unit is in communication with the product chamber; and the liquid sample is rotated in the substrate Under the action of centrifugal force, the reagent chamber flows through the nucleic acid adsorption chamber in turn, and flows to the waste liquid chamber and the product chamber respectively to complete the nucleic acid extraction and purification process. After subsequent detection and analysis of nucleic acid analysis within the unit.

Further, the reagent chamber includes a first reagent chamber, at least one second reagent chamber and a third reagent chamber, and the first reagent chamber is provided with a sample port for adding a liquid sample, and is pre-packaged a cell lysate and a space required for adding a liquid sample; the second reagent chamber is pre-packaged with a washing liquid; and the third reagent chamber is pre-packaged with a nucleic acid that elutes the nucleic acid substance from the nucleic acid adsorption chamber Eluent.

Further, the volume of the liquid sample that can be added to the first reagent chamber is 100 ul to 500 ul.

Further, the reagent chamber is connected to the nucleic acid adsorption chamber through a common or through a respective reagent flow channel; the waste liquid chamber is connected to the nucleic acid adsorption chamber through the waste liquid flow path; the product chamber passes through the product flow The channel is in communication with the nucleic acid adsorption chamber; the waste liquid chamber is further connected to the reagent chamber through the first reagent return air passage; the product chamber is further connected to the reagent chamber through the second reagent return air passage; the nucleic acid The analysis unit is in communication with the product chamber by analyzing the return air passage.

Further, each reagent chamber is provided with a reagent flow channel port and a first reagent return airway port corresponding to the reagent flow channel port, the reagent flow channel port communicates with the reagent flow channel, and the first reagent return airway port and the first reagent return airway Or the second reagent is returned to the airway.

Further, the reagent flow port on each reagent chamber is disposed near the outer side wall of the reagent chamber, and the return air passage port is disposed adjacent to the inner side wall of the reagent chamber.