Novel rapid plasmid extraction device

Abstract

The utility model discloses a novel device for rapidly extracting plasmids, and belongs to the field of biological extraction. Comprising an upper centrifugal tube sleeve and a lower centrifugal tube sleeve which are detachably connected together, the filtering column is arranged in the upper centrifugal pipe sleeve; the centrifugal adsorption column is arranged in the lower centrifugal pipe sleeve; the upper centrifugal tube sleeve is provided with a blocking piece for controlling the upper centrifugal tube sleeve and the lower centrifugal tube sleeve to be communicated and blocked. The plasmid extraction kit has the beneficial effects that the operation is simple, the operation of a plurality of samples is easy to cause confusion of the samples or wrong addition to reduce the efficiency and accuracy of the test, and the tedious operation time and flow of the traditional plasmid extraction kit are greatly solved.

Description

Technical Field

[0001] This application relates to the field of bio-extraction, and more specifically, to a novel device for rapid plasmid extraction. Background Technology

[0002] Bacterial plasmid extraction is a fundamental procedure in molecular biology experiments, aiming to isolate and purify plasmid DNA from bacterial cells. However, this process can be relatively cumbersome, primarily due to the complexity of the experimental steps: plasmid extraction typically involves multiple steps, such as bacterial culture, bacterial collection, bacterial lysis, and the isolation and purification of plasmid DNA. Each step requires careful execution to ensure the quality and integrity of the DNA. Some steps may also require specific reagents and equipment, such as centrifuges and electrophoresis apparatus, which increases the complexity and cost of the experiment.

[0003] To simplify the plasmid extraction process, scientists have developed a variety of commercial plasmid extraction kits. These kits typically provide pre-prepared reagents and optimized procedures, significantly reducing experimental time and improving DNA purity. However, even with kits, the plasmid extraction process remains complex: first, bacterial cells are transferred to centrifuge tubes for collection; then, the bacterial cells are lysed; the lysed cells are added to a filter column, and centrifugation removes the lysed bacterial cells and protein impurities; finally, the collected lysate is added to an adsorption column, where centrifugation adsorbs the plasmids, removing impurity proteins and other organic compounds from the bacteria for plasmid adsorption.

[0004] Traditional plasmid extraction kits require multiple transfers of the lysed resuspension, a relatively cumbersome process that is prone to sample contamination or incorrect addition, reducing experimental efficiency and accuracy. Fully automated multifunctional plasmid extraction systems have changed the situation where medium- to large-volume plasmid extraction mainly relies on manual methods, offering simple and convenient operation. However, their high cost and low throughput limit their widespread use.

[0005] In summary, traditional commercial plasmid extraction kits have relatively cumbersome operation steps, and the operation of multiple samples can easily lead to sample confusion or errors in addition, reducing the efficiency and accuracy of the experiment. Utility Model Content

[0006] The summary section of this application is intended to provide a brief overview of the concepts, which will be described in detail in the detailed description section below. This summary section is not intended to identify key or essential features of the claimed technical solutions, nor is it intended to limit the scope of the claimed technical solutions.

[0007] To address the technical problems mentioned in the background section above, some embodiments of this application provide a novel device for rapid plasmid extraction, comprising: an upper centrifuge tube and a lower centrifuge tube that are detachably connected together; a filter column disposed in the upper centrifuge tube and a centrifugal adsorption column disposed in the lower centrifuge tube; the upper centrifuge tube is provided with a plug for controlling the communication and sealing between the upper centrifuge tube and the lower centrifuge tube.

[0008] Furthermore, the filter column is located at the lower part of the upper centrifuge tube sleeve, and the plug is located at the bottom of the upper centrifuge tube sleeve.

[0009] Furthermore, the centrifugal adsorption column is located below the upper centrifugal sleeve; the bottom of the lower centrifugal sleeve forms a receiving cavity with the centrifugal adsorption column.

[0010] Furthermore, the upper centrifuge tube sleeve is inserted into the lower centrifuge tube sleeve; a gap is formed between the outer wall of the upper centrifuge tube sleeve and the inner wall of the lower centrifuge tube sleeve.

[0011] Furthermore, the plug is a Luer plug.

[0012] Furthermore, a centrifuge tube cap is provided on the upper part of the upper centrifuge tube sleeve, and the centrifuge tube cap is used to seal the upper part of the upper centrifuge tube sleeve.

[0013] The beneficial effects of this application are as follows: bacterial cells can be obtained directly from the bottom tube of the filter column after centrifugation; bacterial cell lysis and neutralization can be performed directly in the upper centrifugation tube; after centrifugation, the lysed cells pass through the filter column and the centrifugal adsorption column in sequence, where the filter column successfully blocks the bacterial cells at the bottom of the filter column; the centrifugal adsorption column can specifically adsorb plasmids, removing impurity proteins and other organic compounds in the bacteria, which can be used to adsorb plasmids to achieve the purpose of plasmid purification; this device is easy to operate, and the operation of multiple samples can easily cause sample confusion or incorrect addition, reducing the efficiency and accuracy of the experiment, greatly solving the cumbersome operation time and process of traditional plasmid extraction kits. Attached Figure Description

[0014] The accompanying drawings, which form part of this application, are used to provide a further understanding of the application and to make other features, objects, and advantages of the application more apparent. The illustrative embodiments and descriptions of this application are used to explain the application and do not constitute an undue limitation of the application.

[0015] Furthermore, throughout the accompanying drawings, the same or similar reference numerals denote the same or similar elements. It should be understood that the drawings are schematic, and the elements are not necessarily drawn to scale.

[0016] In the attached diagram:

[0017] Figure 1 This is an overall schematic diagram based on an embodiment of this application;

[0018] Figure 2 This is a structural schematic diagram of a part of the embodiment, mainly showing... Figure 1 The cross-sectional structure.

[0019] Figure label:

[0020] 1. Centrifuge tube cap; 2. Upper centrifuge tube sleeve; 3. Filter column; 4. Luer plug; 5. Centrifugal adsorption column; 6. Lower centrifuge tube sleeve. Detailed Implementation

[0021] Embodiments of this disclosure will now be described in more detail with reference to the accompanying drawings. While some embodiments of this disclosure are shown in the drawings, it should be understood that this disclosure can be implemented in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of this disclosure. It should be understood that the accompanying drawings and embodiments of this disclosure are for illustrative purposes only and are not intended to limit the scope of protection of this disclosure.

[0022] It should also be noted that, for ease of description, only the parts relevant to the invention are shown in the accompanying drawings. Unless otherwise specified, the embodiments and features described in this disclosure can be combined with each other.

[0023] It should be noted that the concepts of "first" and "second" mentioned in this disclosure are used only to distinguish different devices, modules or units, and are not used to limit the order of functions performed by these devices, modules or units or their interdependencies.

[0024] It should be noted that the terms "a" and "a plurality of" used in this disclosure are illustrative rather than restrictive, and those skilled in the art should understand that, unless otherwise expressly indicated in the context, they should be understood as "one or more".

[0025] This disclosure will now be described in detail with reference to the accompanying drawings and embodiments.

[0026] Reference Figure 1-2 ,

[0027] A novel device for rapid plasmid extraction includes an upper centrifuge tube sleeve 2 and a lower centrifuge tube sleeve 6 that are detachably connected together. The upper centrifuge tube sleeve 2 and the lower centrifuge tube sleeve 6 can be connected by insertion to form a detachable connection, or by a threaded connection. When the upper centrifuge tube sleeve 2 and the lower centrifuge tube sleeve 6 are connected together, the resulting rapid extraction device has two independent chambers. The independent chamber in the upper centrifuge tube sleeve 2 is defined as the upper centrifuge chamber, and the independent chamber in the lower centrifuge tube sleeve 6 is defined as the lower centrifuge chamber.

[0028] The rapid extraction device also includes a filter column 3 disposed in the upper centrifuge tube sleeve 2 and a centrifugal adsorption column 5 disposed in the lower centrifuge tube sleeve 6. The filter column 3 abuts against the inner wall of the upper centrifuge chamber, ensuring that substances in the upper centrifuge chamber must pass through the filter column 3 to move from above to below it. Similarly, the centrifugal adsorption column 5 abuts against the inner wall of the lower centrifuge chamber, thus requiring substances to pass through it for transfer. The core of the filter column 3 is a filter membrane with a specific pore size (such as cellulose or polyethersulfone), which achieves separation through mechanical retention. Large particles (such as cell debris or protein aggregates) are retained on the membrane, while small molecules (such as buffer solutions or small molecule metabolites) or solutions pass through. The core of the centrifugal adsorption column 5 is a filter membrane containing a specific adsorption material (such as silica gel membrane, ion exchange resin, or magnetic beads). Under high-salt, low-pH buffer conditions, target molecules (such as DNA, RNA, and proteins) are adsorbed onto the membrane through hydrogen bonds, ionic interactions, or hydrophobic interactions.

[0029] The upper centrifuge tube sleeve 2 is equipped with a plug for controlling the connection and sealing between the upper centrifuge tube sleeve 2 and the lower centrifuge tube sleeve 6. The upper centrifuge tube is mainly used for adding liquid, and the lower centrifuge tube is mainly used for collecting waste liquid and eluted plasmids. The plug can control the liquid flow in the upper centrifuge tube sleeve 2.

[0030] During use, for example, when collecting bacterial cells before lysis, the plug connects the upper and lower centrifuge chambers. Then, the centrifugal adsorption column 5 in the lower centrifuge chamber is removed, and the bacterial cells are added to the upper centrifuge tube. The device is then placed on a centrifuge for centrifugation, and the bacterial cells are collected below the filter column 3 through centrifugation. Then, the upper centrifuge tube 2 is removed, and the plug is adjusted to disconnect the upper and lower centrifuge chambers. Bacterial resuspension, lysis buffer, and neutralization buffer are added sequentially below the centrifugal adsorption column 5. The upper centrifugal adsorption column 5 is then installed. Centrifugation allows the lysed bacterial cells to pass sequentially through the filter column 3 and the centrifugal adsorption column 5. The plasmids bind to the silica-based centrifugal adsorption column 5 to achieve plasmid purification.

[0031] Specifically, the filter column 3 is located at the lower part of the upper centrifuge tube sleeve 2, and the plug is located at the bottom of the upper centrifuge tube sleeve 2. The centrifugal adsorption column 5 is located below the upper centrifuge tube sleeve 2; the bottom of the lower centrifuge tube sleeve 6 and the centrifugal adsorption column 5 form a receiving cavity. The upper centrifuge tube sleeve 2 is inserted into the lower centrifuge tube sleeve 6; a gap is formed between the outer wall of the upper centrifuge tube sleeve 2 and the inner wall of the lower centrifuge tube sleeve 6. The plug is a Luer plug 4. A centrifuge tube cap 1 is provided at the upper part of the upper centrifuge tube sleeve 2, and the centrifuge tube cap 1 is used to seal the upper part of the upper centrifuge tube sleeve 2.

[0032] This novel plasmid rapid extraction device allows for the direct acquisition of bacterial cells from the bottom tube of the filter column 3 after centrifugation. Adding a Luer plug 4 to the centrifuge tube sleeve 2 enables direct lysis and neutralization of the bacterial cells within the sleeve. After centrifugation, the lysed cells pass sequentially through the filter column 3 and the centrifugal adsorption column 5. The filter column 3 successfully traps the bacterial cells at its bottom. The centrifugal adsorption column 5 specifically adsorbs plasmids, removing impurities, proteins, and other organic compounds from the bacteria, thus purifying the plasmids. This device is easy to operate, addressing the issues of sample confusion or incorrect sample addition during multiple sample operations, which reduce experimental efficiency and accuracy. It significantly solves the problems of time and cumbersome procedures associated with traditional plasmid extraction kits.

[0033] In addition, the specific usage process is as follows: First, remove the Luer plug 4 and centrifugal adsorption column 5 from the upper centrifuge tube, add the bacterial cells to the upper centrifuge tube, and collect the bacterial cells to the bottom of the filter column 3 by centrifugation; add the Luer plug 4, and add the bacterial resuspension, lysis buffer and neutralization buffer in sequence; add the centrifugal adsorption column 5 to the lower centrifuge tube, and centrifuge the lysed bacterial cells to pass through the filter column 3 and centrifugal adsorption column 5 in sequence. The plasmid binds to the silica matrix material centrifugal adsorption column 5 to achieve the purpose of purifying the plasmid.

[0034] The above description is merely a selection of preferred embodiments of this disclosure and an explanation of the technical principles employed. Those skilled in the art should understand that the scope of the invention involved in the embodiments of this disclosure is not limited to technical solutions formed by specific combinations of the above-described technical features, but should also cover other technical solutions formed by arbitrary combinations of the above-described technical features or their equivalents without departing from the above-described inventive concept. For example, technical solutions formed by substituting the above-described features with (but not limited to) technical features with similar functions disclosed in the embodiments of this disclosure.

Claims

1. A novel device for rapid plasmid extraction, characterized in that, include: The upper centrifuge tube sleeve and the lower centrifuge tube sleeve are detachably connected together; a filter column is disposed in the upper centrifuge tube sleeve and a centrifugal adsorption column is disposed in the lower centrifuge tube sleeve; The upper centrifuge sleeve is equipped with a plug for controlling the connection and sealing between the upper centrifuge sleeve and the lower centrifuge sleeve.

2. The novel plasmid rapid extraction device according to claim 1, characterized in that: The filter column is located at the lower part of the upper centrifuge tube sleeve, and the plug is located at the bottom of the upper centrifuge tube sleeve.

3. The novel plasmid rapid extraction device according to claim 1, characterized in that: The centrifugal adsorption column is located below the upper centrifugal tube sleeve; the bottom of the lower centrifugal tube sleeve forms a receiving cavity with the centrifugal adsorption column.

4. The novel plasmid rapid extraction device according to claim 1, characterized in that: The upper centrifuge tube sleeve is inserted into the lower centrifuge tube sleeve; A gap is formed between the outer wall of the upper centrifuge tube and the inner wall of the lower centrifuge tube.

5. The novel plasmid rapid extraction device according to claim 1, characterized in that: The plug is a Luer plug.

6. The novel plasmid rapid extraction device according to claim 1, characterized in that: The upper part of the upper centrifuge tube sleeve is provided with a centrifuge tube cap, which is used to seal the upper part of the upper centrifuge tube sleeve.

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