CN113717837A - Detachable medicinal nucleic acid purification post - Google Patents
Detachable medicinal nucleic acid purification post Download PDFInfo
- Publication number
- CN113717837A CN113717837A CN202110931988.3A CN202110931988A CN113717837A CN 113717837 A CN113717837 A CN 113717837A CN 202110931988 A CN202110931988 A CN 202110931988A CN 113717837 A CN113717837 A CN 113717837A
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- purification column
- nucleic acid
- column
- outer tube
- acid purification
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
- B01D15/1864—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns
- B01D15/1871—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns placed in series
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/22—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the construction of the column
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
- B01D15/3804—Affinity chromatography
- B01D15/3819—Affinity chromatography of the nucleic acid-nucleic acid binding protein type
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D36/00—Filter circuits or combinations of filters with other separating devices
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/101—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
Abstract
The invention discloses a detachable medicinal nucleic acid purification column, which belongs to the technical field of nucleic acid purification and comprises a placing rack and a column body, wherein the column body is erected on the placing rack, and comprises a first purification column, a second purification column and a third purification column; the upper part of the second purifying column is clamped with the lower part of the first purifying column, and the upper part of the third purifying column is clamped with the lower part of the second purifying column; the nucleic acid purification column is divided into a first purification column, a second purification column and a third purification column, so that the nucleic acid purification column is convenient to disassemble and install, continuous operation is realized, excessive biological material transfer steps are avoided, and pollution is prevented; the column body can be erected on the placing rack, so that the use is convenient; the first purification column of the nucleic acid purification column is provided with the filter paper cylinder, so that larger impurities in the biological material which is just crushed can be filtered, and the next separation and purification are facilitated; the filter paper cylinder is arranged in the limiting cylinder, and the limiting cylinder can be detached, so that the filter paper cylinder is convenient to place, clean and replace.
Description
Technical Field
The invention relates to the technical field of nucleic acid purification, in particular to a detachable medicinal nucleic acid purification column.
Background
The extraction and purification of nucleic acid are the basis of molecular biological tests, and the nucleic acid purification method is an important factor influencing the quality of the extracted nucleic acid and is also the key of success or failure of downstream molecular biological tests. Nucleic acid purification in laboratories is commonly performed on nucleic acid purification columns.
The nucleic acid purification column can be used for extracting and refining DNA \ RNA of various materials, and has the characteristics of simple operation, high recovery rate, stable performance and the like. The nucleic acid purification column adopts a silica gel membrane as a specific adsorption material of nucleic acid, does not basically adsorb other biological materials, can ensure to recover DNA \ RNA in a sample to the maximum extent, and simultaneously removes other impurities. The existing nucleic acid purification column is generally selected according to different extraction requirements, the transfer of materials can be involved in the operation process, the operation is careless, the nucleic acid purification result can be affected, and the operation of transferring materials needs to be reduced as much as possible.
Disclosure of Invention
The invention aims to provide a detachable medicinal nucleic acid purification column, which can purify nucleic acid of biological materials through a first purification column, a second purification column and a third purification column which are convenient to detach and install so as to reduce the transfer steps in the purification process.
The purpose of the invention can be realized by the following technical scheme: a detachable medicinal nucleic acid purification column comprises a placing rack and a column body, wherein the column body is erected on the placing rack, and comprises a first purification column, a second purification column and a third purification column; the upper part of the second purifying column is clamped with the lower part of the first purifying column, and the upper part of the third purifying column is clamped with the lower part of the second purifying column;
furthermore, the placing frame comprises a bottom plate and a top plate, wherein four corners of the bottom plate and four corners of the top plate are respectively connected through supporting rods, a frame plate is arranged between the bottom plate and the top plate, and the four corners of the frame plate are respectively fixedly connected with the four supporting rods; the frame plate and the top plate are both provided with notches for erecting the column body;
further, the first purification column comprises a first outer tube, a first collecting pipe is arranged at the bottom of the first outer tube, and a circular expanded annular flange is arranged at the top of the first outer tube; the top end of the first outer pipe is provided with a limiting cylinder, the limiting cylinder comprises a limiting ring, a middle ring and a base, the limiting ring, the middle ring and the base are fixedly connected through three connecting rods which are uniformly arranged in the circumferential direction, and the middle ring is arranged between the limiting ring and the base;
the outer diameters of the centering ring and the base are smaller than the inner diameter of the first outer tube by the difference of D1, and D1 is 1.5-4 mm; the outer diameter of the limiting ring is larger than the inner diameter of the first outer pipe, and the limiting ring is erected on the annular flange of the first outer pipe; a filter paper cylinder is arranged in the limiting cylinder and is formed by winding rectangular filter paper; the base is provided with an annular limiting groove corresponding to the filter paper cylinder, and one end of the filter paper cylinder is clamped in the limiting groove;
further, the second purification column comprises a second outer tube; the upper end of the second outer pipe is provided with a butterfly plate, the lower end of the second outer pipe is provided with a clamping pipe, and the lower end of the clamping pipe is provided with a second collecting pipe; a first filter plate is arranged at the bottom of the second outer pipe and is fixed through a first pressing ring; the clamping pipe and the second collecting pipe are provided with air grooves for balancing air pressure;
the inner diameters of the second outer pipe and the first collecting pipe are the same, so that clamping fixation is facilitated; the second outer tube is also provided with a piston rod in a matching way, the piston rod comprises a push rod, and the bottom of the push rod is fixedly provided with a piston head; the diameter of the piston head is larger than the inner diameter of the second outer tube by the difference D2, and D2 is 0.5-0.8 mm;
further, the third purifying column comprises a third outer tube, an inner tube is sleeved in the third outer tube, and the inner diameter of the third outer tube is the same as the outer diameter of the inner tube; the lower end of the inner pipe is provided with a third collecting pipe, and the upper end of the inner pipe is provided with a lantern ring; the bottom of the inner tube is provided with a second filter plate and is fixed by a second pressing ring; when the second purifying column is connected with the third purifying column, the clamping pipe is clamped with the lantern ring, and the second collecting pipe extends into the inner pipe; the upper end of the third outer tube is connected with a plug through a soft belt, when the third purification column is arranged in a centrifuge for operation, the inner tube is plugged to prevent liquid leakage, and a protective shell is clamped outside the plug to prevent the column body from being polluted when other operation steps are carried out;
the first pressing ring and the second pressing ring do not adsorb most of biomolecules, and the second filter plate can adsorb nucleic acid molecules; the detachable medicinal nucleic acid purification column can be provided with different specifications according to the requirement of nucleic acid purification.
The invention has the beneficial effects that:
1. the nucleic acid purification column is divided into a first purification column, a second purification column and a third purification column, so that the nucleic acid purification column is convenient to disassemble and install, continuous operation is realized, excessive biological material transfer steps are avoided, and pollution is prevented; the column body can be erect to the rack, facilitates the use.
2. The first purification column of the nucleic acid purification column is provided with the filter paper cylinder, so that larger impurities in the biological material which is just crushed can be filtered, and the next separation and purification are facilitated; the filter paper cylinder is arranged in the limiting cylinder, and the limiting cylinder can be detached, so that the filter paper cylinder is convenient to place, clean and replace.
3. The second outer tube can collect the biological material purified by the first purifying column, at this time, the column body is taken down from the placing frame, the first purifying column is disassembled, and the biological material in the first outer tube can be further purified and sent into the inner tube by matching with the piston rod; the clamping pipe and the collecting pipe are provided with air grooves, so that air pressure can be balanced when the piston rod is pushed.
4. After the operation of the second purification column part is completed, when the third purification column is arranged in the centrifuge operation, the inner tube can be plugged by the plug to prevent liquid leakage, and the plug is externally connected with a protective shell in a clamping manner to prevent the cylinder from being polluted when other operation steps are carried out.
Drawings
The invention will be further described with reference to the accompanying drawings.
FIG. 1 is a schematic structural view of the present invention;
figure 2 is a schematic view of the rack of the present invention;
FIG. 3 is an exploded view of a first purification column according to the present invention;
FIG. 4 is a schematic view of a spacing cylinder of the present invention;
FIG. 5 is a schematic view of a second outer tube section of the present invention;
FIG. 6 is a schematic view of the second and third purification columns of the present invention;
FIG. 7 is a schematic view of a third purification column of the present invention;
FIG. 8 is an exploded view of a third purification column according to the present invention;
fig. 9 is a schematic view of a piston rod of the present invention.
In the figure: 1. placing a rack; 2. a first purification column; 3. a second purification column; 4. a third purification column; 11. a base plate; 12. a frame plate; 13. a top plate; 14. a support bar; 21. a first outer tube; 22. a first header; 23. a limiting cylinder; 24. a filter paper cartridge; 231. a limiting ring; 232. a neutral ring; 233. a base; 234. a connecting rod; 235. a limiting groove; 31. a second outer tube; 32. a butterfly plate; 33. a first filter plate; 34. a first clamping ring; 35. clamping the connecting pipe; 36. a second header; 37. an air tank; 38. a piston rod; 381. a push rod; 382. a piston head; 41. a third outer tube; 42. an inner tube; 43. a third header pipe; 44. a collar; 45. a second filter plate; 46. a second clamping ring; 47. a plug; 48. a protective shell.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-9, the present invention is a detachable pharmaceutical nucleic acid purification column, comprising a rack 1 and a column body, wherein the column body is erected on the rack 1, and comprises a first purification column 2, a second purification column 3 and a third purification column 4; the upper part of the second purifying column 3 is clamped with the lower part of the first purifying column 2, and the upper part of the third purifying column 4 is clamped with the lower part of the second purifying column 3; the placing frame 1 comprises a bottom plate 11 and a top plate 13, wherein four corners of the bottom plate 11 and four corners of the top plate 13 are respectively connected through supporting rods 14, a frame plate 12 is arranged between the bottom plate 11 and the top plate 13, and the four corners of the frame plate 12 are respectively fixedly connected with the four supporting rods 14; the frame plate 12 and the top plate 13 are both provided with notches for erecting columns;
the first purification column 2 comprises a first outer tube 21, a first collecting pipe 22 is arranged at the bottom of the first outer tube 21, and a circular expanded annular flange is arranged at the top of the first outer tube 21; the top end of the first outer tube 21 is provided with a limiting cylinder 23, the limiting cylinder 23 comprises a limiting ring 231, a middle position ring 232 and a base 233, the limiting ring 231, the middle position ring 232 and the base 233 are fixedly connected through three connecting rods 234 uniformly arranged in the circumferential direction, and the middle position ring 232 is arranged between the limiting ring 231 and the base 233; the outer diameters of the centering ring 232 and the base 233 are smaller than the inner diameter of the first outer tube 21; the outer diameter of the retainer ring 231 is larger than the inner diameter of the first outer tube 21, and is mounted on the annular flange of the first outer tube 21; a filter paper cylinder 24 is arranged in the limiting cylinder 23, and the filter paper cylinder 24 is formed by winding rectangular filter paper; the base 233 is provided with a limit groove 235 corresponding to the filter paper cylinder 24, and one end of the filter paper cylinder 24 is clamped in the limit groove 235;
the second purification column 3 comprises a second outer tube 31; a butterfly plate 32 is arranged at the upper end of the second outer pipe 31, a clamping pipe 35 is arranged at the lower end of the second outer pipe, and a second collecting pipe 36 is arranged at the lower end of the clamping pipe 35; a first filter plate 33 is arranged at the bottom of the second outer tube 31, and the first filter plate 33 is fixed by a first pressing ring 34; the clamping pipe 35 and the second collecting pipe 36 are provided with air grooves 37 for balancing air pressure; the inner diameters of the second outer pipes 31 and the first collecting pipes 22 are the same, so that clamping fixation is facilitated; the second outer tube 31 is further provided with a piston rod 38 in a matching manner, the piston rod 38 comprises a push rod 381, and a piston head 382 is fixedly arranged at the bottom of the push rod 381; the piston head 382 has a diameter greater than the inner diameter of the second outer tube 31;
the third purification column 4 comprises a third outer tube 41, an inner tube 42 is sleeved in the third outer tube 41, and the inner diameter of the third outer tube 41 is the same as the outer diameter of the inner tube 42; the lower end of the inner pipe 42 is provided with a third collecting pipe 43, and the upper end is provided with a lantern ring 44; the bottom of the inner tube 42 is provided with a second filter plate 45 and fixed by a second pressing ring 46; when the second purifying column 3 is connected with the third purifying column 4, the clamping pipe 35 is clamped with the lantern ring 44, and the second collecting pipe 36 extends into the inner pipe 42; the upper end of the third outer tube 41 is connected with a plug 47, when the third purification column 4 is arranged in the centrifuge for operation, the inner tube 42 is plugged to prevent liquid leakage, and the plug 47 is also clamped with a protective shell 48 to prevent the column body from being polluted when other operation steps are carried out.
The working principle of the invention is as follows: firstly, assembling a first purifying column 2, a second purifying column 3 and a third purifying column 4 into a complete column body, erecting the column body on a placing frame 1, arranging a circular unfolded annular flange at the top of a first outer tube 21 and erecting the annular flange on a top plate 13, and erecting a butterfly plate 32 on a frame plate 12;
crushing cell biological materials, adding the crushed cell biological materials into the first purification column 2, filtering larger impurities by using a filter paper cylinder 24, and allowing the crude extracted biological materials to flow into a second outer tube 31 along a first collecting pipe 22; taking down the column body, then taking down the first purification column 2 part, plugging the piston rod 38 into the second outer tube 31, pressing the push rod 381, filtering the crude extracted biological material by the first filter plate 33 to obtain a middle extract, and allowing the middle extract to flow into the inner tube 42 along the second collecting pipe 36; taking off the second purification column 3 part, taking off the protective shell 48, plugging the plug 47 into the inner tube 42, and placing the third purification column 4 in a centrifuge for centrifugation; the nucleic acid molecules in the middle extract are adsorbed by the second filter plate 45 and further separated and purified, and the residual liquid enters the bottom of the third outer tube 41 through centrifugation.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "left", "right", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are only for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation and a specific orientation configuration and operation, and thus, should not be construed as limiting the present invention. Furthermore, "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and the like are to be construed broadly and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be directly connected or indirectly connected through an intermediate member, or they may be connected through two or more elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
While one embodiment of the present invention has been described in detail, the description is only a preferred embodiment of the present invention and should not be taken as limiting the scope of the invention. All equivalent changes and modifications made within the scope of the present invention shall fall within the scope of the present invention.
Claims (10)
1. A detachable medicinal nucleic acid purification column comprises a placing rack (1) and a column body, wherein the column body is erected on the placing rack (1), and is characterized in that the column body comprises a first purification column (2), a second purification column (3) and a third purification column (4); the first purifying column (2), the second purifying column (3) and the third purifying column (4) are clamped in sequence.
2. The removable medicinal nucleic acid purification column according to claim 1, wherein the rack (1) comprises a bottom plate (11) and a top plate (13), four corners of the bottom plate (11) and the top plate (13) are respectively connected through support rods (14), and a frame plate (12) is connected between the bottom plate (11) and the top plate (13) through the support rods (14); the frame plate (12) and the top plate (13) are both provided with notches.
3. The removable medicinal nucleic acid purification column according to claim 1, wherein the first purification column (2) comprises a first outer tube (21), a first collecting pipe (22) is arranged at the bottom of the first outer tube (21), and an annular flange is arranged at the top of the first outer tube (21); the top end of the first outer tube (21) is provided with a limiting cylinder (23), and the limiting cylinder (23) comprises a limiting ring (231), a middle ring (232) and a base (233) which are fixedly connected through a plurality of connecting rods (234) which are uniformly arranged in the circumferential direction.
4. The removable pharmaceutical nucleic acid purification column of claim 3, wherein the outer diameter of the centering ring (232) and the base (233) is smaller than the inner diameter of the first outer tube (21); the outer diameter of the limiting ring (231) is larger than the inner diameter of the first outer pipe (21).
5. The removable medicinal nucleic acid purification column of claim 3, wherein the limiting cylinder (23) is provided with a filter paper cylinder (24); the base (233) is provided with a limit groove (235), and one end of the filter paper cylinder (24) is clamped in the limit groove (235).
6. The removable pharmaceutical nucleic acid purification column according to claim 1, wherein the second purification column (3) comprises a second outer tube (31); a butterfly plate (32) is arranged at the upper end of the second outer pipe (31), a clamping pipe (35) is arranged at the lower end of the second outer pipe, and a second collecting pipe (36) is arranged at the lower end of the clamping pipe (35); the bottom of the second outer tube (31) is provided with a first filter plate (33), and the first filter plate (33) is fixed through a first pressing ring (34).
7. The removable pharmaceutical nucleic acid purification column according to claim 6, wherein the clamping tube (35) and the second collecting tube (36) are provided with air grooves (37).
8. The removable pharmaceutical nucleic acid purification column according to claim 6, wherein the second outer tube (31) is provided with a piston rod (38), the piston rod (38) comprises a push rod (381), and the bottom of the push rod (381) is fixedly provided with a piston head (382); the piston head (382) has a diameter greater than the inner diameter of the second outer tube (31).
9. The removable pharmaceutical nucleic acid purification column according to claim 1, wherein the third purification column (4) comprises a third outer tube (41), and an inner tube (42) is sleeved inside the third outer tube (41); the lower end of the inner pipe (42) is provided with a third collecting pipe (43), and the upper end is provided with a lantern ring (44); the bottom of the inner tube (42) is provided with a second filter plate (45) and fixed by a second pressing ring (46).
10. The removable pharmaceutical nucleic acid purification column according to claim 9, wherein the upper end of the third outer tube (41) is connected with a plug (47), and a protective shell (48) is clamped outside the plug (47).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110931988.3A CN113717837B (en) | 2021-08-13 | 2021-08-13 | Detachable medicinal nucleic acid purification column |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110931988.3A CN113717837B (en) | 2021-08-13 | 2021-08-13 | Detachable medicinal nucleic acid purification column |
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| CN113717837A true CN113717837A (en) | 2021-11-30 |
| CN113717837B CN113717837B (en) | 2023-10-03 |
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| CN211035888U (en) * | 2019-10-16 | 2020-07-17 | 南京君华基因科技有限公司 | Free nucleic acid piece-rate system of pressurization formula |
| CN211664984U (en) * | 2019-12-05 | 2020-10-13 | 南京翼飞雪生物科技有限公司 | Purification device is used in biological antibody production |
| CN110904095A (en) * | 2019-12-12 | 2020-03-24 | 杭州倍强医药科技有限公司 | Method for improving small fragment nucleic acid purification yield |
| CN212293531U (en) * | 2020-06-19 | 2021-01-05 | 上海快灵生物科技有限公司 | Nucleic acid purification system |
| CN213232220U (en) * | 2020-09-02 | 2021-05-18 | 济凡生物科技(常州)有限公司 | A novel liquefaction post device for blood clot nucleic acid draws |
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