CN113930318B - Nucleic acid extraction consumables and method for extracting nucleic acid using the same - Google Patents
Nucleic acid extraction consumables and method for extracting nucleic acid using the same Download PDFInfo
- Publication number
- CN113930318B CN113930318B CN202010668889.6A CN202010668889A CN113930318B CN 113930318 B CN113930318 B CN 113930318B CN 202010668889 A CN202010668889 A CN 202010668889A CN 113930318 B CN113930318 B CN 113930318B
- Authority
- CN
- China
- Prior art keywords
- piston
- liquid
- nucleic acid
- outer cylinder
- filter paper
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 75
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 75
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 75
- 238000000605 extraction Methods 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 166
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 claims abstract description 67
- 238000002347 injection Methods 0.000 claims abstract description 44
- 239000007924 injection Substances 0.000 claims abstract description 44
- 230000007246 mechanism Effects 0.000 claims abstract description 22
- 238000007789 sealing Methods 0.000 claims description 70
- 239000003480 eluent Substances 0.000 claims description 32
- 239000002699 waste material Substances 0.000 claims description 24
- 238000005336 cracking Methods 0.000 claims description 20
- 239000012488 sample solution Substances 0.000 claims description 20
- 238000000197 pyrolysis Methods 0.000 claims description 18
- 230000009471 action Effects 0.000 claims description 15
- 230000005484 gravity Effects 0.000 claims description 14
- 238000007599 discharging Methods 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 11
- 238000003825 pressing Methods 0.000 claims description 9
- 230000008569 process Effects 0.000 claims description 9
- 239000000523 sample Substances 0.000 claims description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000012864 cross contamination Methods 0.000 abstract description 3
- 238000010586 diagram Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 3
- 239000004743 Polypropylene Substances 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000001125 extrusion Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- -1 polypropylene Polymers 0.000 description 2
- 229920001155 polypropylene Polymers 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 1
- 206010011409 Cross infection Diseases 0.000 description 1
- 206010029803 Nosocomial infection Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
本发明公开了一种核酸提取耗材及使用其提取核酸的方法，属于核酸提取技术领域。本发明包括外筒以及活塞，所述活塞与外筒配合且能够沿着外筒上下移动；所述活塞内部中空，所述活塞中空部分的上口为注液口；所述活塞的中空部分构成流道，所述流道内设有止回机构；所述外筒内底部设置若干层硅酸盐滤纸；所述外筒底部设置出液口。本发明为全新的核酸提取工具。这种新型耗材在保证提取过程完全密闭、无交叉污染风险的前提下，通过活塞上移、下移轻松实现核酸提取。
The present invention discloses a nucleic acid extraction consumable and a method for extracting nucleic acid using the same, and belongs to the technical field of nucleic acid extraction. The present invention comprises an outer cylinder and a piston, wherein the piston cooperates with the outer cylinder and can move up and down along the outer cylinder; the interior of the piston is hollow, and the upper opening of the hollow part of the piston is a liquid injection port; the hollow part of the piston constitutes a flow channel, and a check mechanism is provided in the flow channel; several layers of silicate filter paper are provided at the bottom of the outer cylinder; and a liquid outlet is provided at the bottom of the outer cylinder. The present invention is a brand-new nucleic acid extraction tool. This new consumable can easily realize nucleic acid extraction by moving the piston up and down, while ensuring that the extraction process is completely closed and there is no risk of cross contamination.
Description
Technical Field
The invention relates to the technical field of accounting extraction, in particular to a nucleic acid extraction consumable and a method for extracting nucleic acid by using the same.
Background
Nucleic acid extraction is widely used in the fields of molecular biology, biochemistry, food safety, pathogenic microorganism molecular diagnosis, genomics and the like. In recent years, with serious threat of infectious diseases, instant detection based on molecular diagnosis technology is rapidly developed. These assays all require the extraction of the nucleic acid of the sample prior to detection.
In the current biochemical laboratory, there are two main methods of nucleic acid extraction, namely, a filter column method and a magnetic bead method. The extraction rate of the filter column method is higher, but the large-scale automatic extraction is difficult to achieve due to the requirement of a centrifugal machine, and the operation is complex, so that the time and the labor are wasted; meanwhile, the requirements on experimental environment are strict; while the magnetic bead method can be matched with a nucleic acid extractor for automatic extraction, the method has the defects of low extraction rate, cross contamination and the like, and the extracted nucleic acid sample has a plurality of problems.
Therefore, it is important to improve the nucleic acid extraction rate and the nucleic acid extraction efficiency while ensuring no cross infection.
Disclosure of Invention
In order to make up for the defects of the prior art, the invention provides a nucleic acid extraction consumable and a method for extracting nucleic acid by using the same. The nucleic acid extraction consumable of the invention has the advantages of totally-enclosed design, high extraction efficiency and automation.
The technical scheme of the invention is as follows:
The nucleic acid extraction consumable comprises an outer cylinder and a piston, wherein the piston is matched with the outer cylinder and can move up and down along the outer cylinder; the piston is hollow, and the upper opening of the hollow part of the piston is a liquid injection opening; the hollow part of the piston forms a flow passage, and a non-return mechanism is arranged in the flow passage; a plurality of layers of silicate filter paper are arranged at the inner bottom of the outer cylinder; the bottom of the outer cylinder is provided with a liquid outlet.
A method for extracting nucleic acid using the nucleic acid extraction consumable, comprising the steps of:
1) Injecting sample solution into the outer cylinder through the liquid injection port, wherein the sample solution enters silicate filter paper;
2) Injecting the pyrolysis liquid into the outer cylinder from the liquid injection port, and allowing the pyrolysis liquid to enter the silicate filter paper
3) After the cracking is finished, the nucleic acid molecules are adsorbed by the silicon dioxide groups on the silicate filter paper, the piston is moved downwards, the piston is continuously extruded downwards after contacting the silicate filter paper, the waste liquid of the cracking liquid is discharged from the liquid outlet at the bottom, and the piston returns to the initial position after the waste liquid of the cracking liquid is discharged completely; the non-return mechanism prevents the solution from flowing back in the process of returning the piston to the initial position;
4) Injecting rinsing liquid into the outer cylinder through the liquid injection port, wherein the rinsing liquid enters silicate filter paper;
5) After standing for a period of time, moving the piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the waste liquid of the rinsing liquid at a liquid outlet at the bottom, and returning the piston to the initial position after the waste liquid of the rinsing liquid is discharged completely; the non-return mechanism prevents the solution from flowing back in the process of returning the piston to the initial position;
6) Repeating step 4) and step 5) for several times;
7) Injecting eluent into the outer cylinder from the liquid injection port, and allowing the eluent to enter silicate filter paper;
8) Standing for a period of time, eluting the nucleic acid molecules adsorbed on the silicate filter paper, moving the piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the eluent and the eluted nucleic acid molecules at the bottom liquid outlet, and dripping into a sample collector; after the eluent is discharged, the piston returns to the initial position; the non-return mechanism prevents back flow of the solution during return of the piston to the initial position.
Preferably, a sealing ring is arranged between the piston and the inner wall of the outer cylinder.
Further, the sealing ring is a rubber O-shaped ring.
Preferably, an opening is arranged at the upper part of the outer cylinder, and the piston comprises an upper piston and a lower piston; the upper piston can slide up and down along the upper opening of the outer cylinder; the lower piston can slide up and down along the inner wall of the outer cylinder and is in sealing connection with the inner wall of the outer cylinder; the upper piston and the lower piston are connected into a whole; the upper piston and the lower piston are hollow.
Further, the runner at least comprises a first sub-runner and a second sub-runner from top to bottom, and the inner diameter of the second sub-runner is larger than that of the first sub-runner.
Further, the check mechanism comprises a spring and a sealing ball fixed on the spring, and the bottom of the spring is fixed in the flow passage; the original position of the sealing ball is the lowest part of the first sub-runner with the top sealed.
A method for extracting nucleic acid using the nucleic acid extraction consumable, comprising the steps of:
1) Injecting sample solution into the outer cylinder through the liquid injection port, pressing the spring under the action of gravity of the sample solution, allowing the sealing ball to move downwards, allowing the sample solution to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then entering silicate filter paper, allowing the spring to rebound after the sample solution passes through the silicate filter paper, and resetting the sealing ball;
2) Injecting a pyrolysis liquid into the outer cylinder from the liquid injection port, pressing the spring under the action of gravity of the pyrolysis liquid, allowing the sealing ball to move downwards, allowing the pyrolysis liquid to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then allowing the pyrolysis liquid to enter silicate filter paper, allowing the spring to rebound after the pyrolysis liquid passes through, and allowing the sealing ball to reset;
3) After the cracking is finished, the nucleic acid molecules are adsorbed by the silicon dioxide groups on the silicate filter paper, the upper piston and the lower piston are moved downwards, the lower piston is continuously extruded downwards after the nucleic acid molecules contact the silicate filter paper, the waste liquid of the cracking liquid is discharged from the liquid outlet at the bottom, and after the waste liquid of the cracking liquid is discharged, the upper piston and the lower piston are moved upwards to return to the initial position;
4) Injecting rinsing liquid into the outer cylinder from the liquid injection port, pressing the spring under the action of the gravity of the rinsing liquid, allowing the sealing ball to move downwards, allowing the rinsing liquid to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then allowing the rinsing liquid to enter silicate filter paper, allowing the spring to rebound after the rinsing liquid passes through the sealing ball, and allowing the sealing ball to reset;
5) After standing for a period of time, moving the upper piston and the lower piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the waste liquid of the rinsing liquid at a liquid outlet at the bottom, and moving the upper piston and the lower piston upwards to return to the initial position after the waste liquid of the rinsing liquid is discharged completely;
6) Repeating step 4) and step 5) for several times;
7) Injecting eluent into the outer cylinder from the liquid injection port, under the action of gravity of the eluent, pressing the spring, moving the sealing ball downwards, enabling the eluent to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then entering silicate filter paper, and returning the sealing ball after the eluent passes through the sealing ball;
8) Standing for a period of time, after the nucleic acid molecules adsorbed on the silicate filter paper are eluted, moving the upper piston and the lower piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the eluent and the eluted nucleic acid molecules at a liquid outlet at the bottom, and dripping the eluent and the eluted nucleic acid molecules into a sample collector; after the eluent is discharged, the upper piston and the lower piston are moved upwards to return to the initial positions.
Further, the flow channel consists of a first flow dividing channel, a second flow dividing channel, a third flow dividing channel and a fourth flow dividing channel which are distributed from top to bottom; the inner diameter of the third sub-runner is smaller than that of the second sub-runner; the inner diameter of the fourth sub-runner is smaller than that of the third sub-runner. The runner is designed to be in a ladder shape, and the bottom of the spring is fixed on a platform of the lower piston, so that the spring can be conveniently and stably fixed.
As a preferable scheme, a first sealing ring is arranged between the upper piston and the lower piston; and a second sealing ring is arranged between the lower piston and the inner wall of the outer cylinder. Two sealing rings are arranged to ensure complete sealing of the internal environment.
Further, the first sealing ring and the second sealing ring are rubber O-shaped rings.
As a preferable scheme, the non-return mechanism comprises a support column fixed in the runner, a spring is sleeved outside the support column, and a sealing head is connected to the support column in a sliding manner; the sliding connection part of the closing head and the supporting column is positioned above the spring.
Preferably, the upper piston is detachably connected with the lower piston.
Further, the bottom outer wall of the upper piston is in threaded connection with the upper inner wall of the lower piston.
As a preferable scheme, 5-8 layers of silicate filter paper are arranged. According to the sample solution amount, the nucleic acid molecules in the sample can be fully extracted by arranging 5-8 layers of silicate filter paper.
Further, 6 layers of silicate filter paper were provided.
Preferably, the outer cylinder, the upper piston and the lower piston are all made of polypropylene materials.
The beneficial effects of the invention are as follows:
1. the nucleic acid extraction consumable of the invention is a brand-new nucleic acid extraction tool, and the novel consumable can easily realize nucleic acid extraction by upward and downward movement of the piston on the premise of ensuring that the extraction process is completely closed and no cross contamination risk exists.
2. The nucleic acid extraction consumable can adopt the motor to drive the piston to extrude with high precision, and the silicate filter paper with high extraction rate is adopted, so that an automatic extraction flow is realized, the extracted nucleic acid is ensured to have higher purity and concentration, and the labor consumption in the experimental process is greatly reduced.
3. A sealing ball is arranged in the flow channel, and a spring is arranged below the sealing ball, so that the sealing ball is not sealed to the flow channel when liquid needs to be injected, and the normal injection of the liquid is ensured; after the liquid is injected, the spring rebounds, and the sealing ball re-seals the flow channel, so that the situation that liquid flows back in the process of extrusion and resetting of the piston is avoided.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that other drawings can be obtained according to these drawings without inventive faculty for a person skilled in the art.
FIG. 1 is a schematic diagram of the nucleic acid isolation consumable of example 1 of the present invention;
FIG. 2 is a schematic diagram showing the initial state of nucleic acid extraction using the consumable of example 1;
FIG. 3 is a schematic view showing the state of the closing ball pressing down the liquid into the liquid storage region by the gravity action of the liquid during the nucleic acid extraction using the consumable of example 1;
FIG. 4 is a schematic diagram showing the state of the upper piston and the lower piston moving downward to squeeze the silicate filter paper during the nucleic acid extraction using the consumable of example 1;
FIG. 5 is a schematic diagram of the structure of the nucleic acid extracting consumable of the present invention;
FIG. 6 is a schematic diagram of another check mechanism in a flow channel of a nucleic acid isolation consumable of the present invention.
Detailed Description
The invention is further explained below with reference to the drawings.
In the description of the present invention, the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "vertical", "horizontal", etc. refer to the orientation or positional relationship based on that shown in the drawings, and are merely for the purpose of describing the present invention and do not require that the present invention must be constructed or operated in a specific orientation, and thus should not be construed as limiting the present invention. "connected" and "connected" in the present invention are to be understood broadly, and may be, for example, connected or detachably connected; the connection may be direct or indirect through intermediate members, and the specific meaning of the terms may be understood in detail by those skilled in the art.
Example 1
As shown in FIG. 1, the nucleic acid extraction consumable of the present invention comprises an outer cylinder 7, wherein the upper part of the outer cylinder 7 is provided with a circular opening, and the lower part is provided with a liquid outlet 8.
Six layers of silicate filter papers 9 are arranged above the liquid outlet 8 in the outer barrel 7, and the silicate filter papers 9 are loosely arranged.
The circular opening of urceolus 7 upper portion department is equipped with upper piston 3, and upper piston 3 inside cavity, according to inside diameter, upper piston 3 divide into thin section and thick section, and upper piston 3's thin section outer wall meets with the circular opening of urceolus 7 upper portion to can follow the circular opening of urceolus 7 upper portion and slide from top to bottom. The lower part of the upper piston 3 has a reduced wall thickness, and the thinner part is provided with an external thread by which the upper piston 3 is screwed with the lower piston 5.
The outer wall of the lower piston 5 is connected with the inner wall of the outer cylinder 7, and the lower piston 5 can slide up and down along the inner wall of the outer cylinder 7. The inner wall of the lower piston 5 is arranged in a stepped manner, the upper wall of the lower piston 5 is thinnest, then the thickness is sequentially increased twice, the thinnest part of the lower piston 5 is provided with internal threads, and the internal threads are matched and connected with external threads on the upper piston 3.
The top of the position with the largest wall thickness of the lower piston 5 is fixed with a spring 11, and a sealing ball 12 is fixed on the spring 11. When the spring 11 is not compressed, the top of the closing ball 12 closes the outlet of the thin section of the upper piston 3.
In order to ensure the tightness of the consumable, a first sealing ring 4 is arranged above the joint of the upper piston 3 and the lower piston 5; a second sealing ring 6 is arranged between the lower piston 5 and the inner wall of the outer cylinder 7. In this embodiment, the first seal ring 4 and the second seal ring 6 are rubber O-rings.
The outer cylinder 7, the upper piston 3 and the lower piston 5 are all made of polypropylene materials, the hollow parts of the upper piston 3 and the lower piston 5 form a runner 2, and the top of the runner 2 is provided with a liquid injection port 1.
The flow channel 2 consists of a first flow dividing channel, a second flow dividing channel, a third flow dividing channel and a fourth flow dividing channel from top to bottom due to the influence of the wall thickness or the inner diameter of the upper piston 3 and the lower piston 5; the inner diameter of the first sub-runner is smaller than that of the second sub-runner, and the sealing ball 12 is arranged between the first sub-runner and the second sub-runner; the inner diameter of the third sub-runner is smaller than that of the second sub-runner; the inner diameter of the fourth sub-runner is smaller than that of the third sub-runner.
The method for extracting nucleic acid by using the nucleic acid extraction consumable comprises the following steps:
1) The injection gun head matched with the liquid injection port 1 is inserted into the consumable along the liquid injection port 1, after the consumable is fixed, the piston 3 is pulled up, the part of the upper piston 3 positioned in the outer cylinder 7 is contacted with the uppermost part of the inner wall of the outer cylinder 7 (a specific position is shown as figure 2), and the tightness of the interior of the consumable is ensured;
2) The injection gun head injects sample solution into the outer barrel 7 from the liquid injection port 1, the spring 11 is pressed under the action of gravity of the sample solution, the sealing ball 12 moves downwards, the sample solution flows into the liquid storage area 10 (shown in figure 3) through a gap between the sealing ball 12 and the upper piston 3, then enters the silicate filter paper 9, and after the sample solution passes through, the spring 11 rebounds, and the sealing ball 12 resets;
3) The injection gun head injects the pyrolysis liquid into the outer barrel 7 from the liquid injection port 1, under the action of gravity of the pyrolysis liquid, the spring 11 is pressed, the sealing ball 12 moves downwards, the pyrolysis liquid flows into the liquid storage area 10 (shown in figure 3) through a gap between the sealing ball 12 and the upper piston 3, then enters the silicate filter paper 9, and after the pyrolysis liquid passes through, the spring 11 rebounds, and the sealing ball 12 resets;
4) After the cracking is finished, the nucleic acid molecules are adsorbed by the silicon dioxide groups on the silicate filter paper 9, the upper piston 3 and the lower piston 5 are moved downwards, the lower piston 5 is continuously extruded downwards after the contact of the silicate filter paper 9, the waste liquid of the cracking liquid is discharged from the bottom liquid outlet 8 (as shown in figure 4), and after the waste liquid of the cracking liquid is discharged, the upper piston 3 and the lower piston 5 are moved upwards to return to the initial positions;
5) The injection gun head injects rinsing liquid into the outer cylinder 7 from the liquid injection port 1, under the action of the gravity of the rinsing liquid, the spring 11 is pressed, the sealing ball 12 moves downwards, the rinsing liquid flows into the liquid storage area 10 (shown in figure 3) through a gap between the sealing ball 12 and the upper piston 3, then enters the silicate filter paper 9, after the rinsing liquid passes through, the spring 11 rebounds, and the sealing ball 12 resets;
6) After standing for a period of time, the upper piston 3 and the lower piston 5 are moved downwards, the lower piston is continuously extruded downwards after contacting the silicate filter paper 9 (as shown in fig. 4), the waste liquid of the rinsing liquid is discharged from the liquid outlet 8 at the bottom, and after the waste liquid of the rinsing liquid is discharged, the upper piston 3 and the lower piston 5 are moved upwards to return to the initial position;
7) Repeating step 5) and step 6) for a plurality of times;
8) The injection gun head injects eluent into the outer cylinder 7 from the injection port 1, under the action of the gravity of the eluent, the spring 11 is pressed, the sealing ball 12 moves downwards, the eluent flows into the liquid storage area 10 (shown in figure 3) through a gap between the sealing ball 12 and the upper piston 3, then enters the silicate filter paper 9, and after the eluent passes, the spring 11 rebounds, and the sealing ball 12 resets;
9) Standing for a period of time, eluting the nucleic acid molecules to be adsorbed on the silicate filter paper 9, then moving the upper piston 3 and the lower piston 5 downwards, continuously extruding downwards after contacting the silicate filter paper 9, discharging the eluent and the eluted nucleic acid molecules at the bottom liquid outlet 8 (as shown in fig. 4), and dripping into a sample collector; after the eluent is discharged, the upper piston 3 and the lower piston 5 are moved upwards to return to the initial positions.
When in actual use, the liquid injection port is matched with the injection gun head, so that the injection gun head is fixed with the consumable material of the invention, and further, the injection of sample solution and the like is realized. The form of the injection gun head and the injection port can also be replaced.
In other embodiments, the flow passage can be separately arranged in the hollow part of the piston and fixedly connected with the upper piston and the lower piston.
In other embodiments, the fixing mode of the spring can be changed as long as the sealing effect of the sealing ball on the flow channel can be realized, so that the situation that liquid flows back in the process of extrusion and resetting of the piston is avoided.
Example 2
As shown in fig. 5, a nucleic acid extraction consumable comprises an outer cylinder 7 and a piston 14, wherein the piston 14 is matched with the outer cylinder 7 (the outer wall of the piston 14 is sealed with the inner wall of the outer cylinder 7 (similar to the matching way of the outer wall of the piston of a syringe and the inner wall of the syringe) or sealed by a sealing ring) and can move up and down along the outer cylinder 7; the interior of the piston 14 is hollow, and the upper opening of the hollow part of the piston 14 is a liquid injection opening; the hollow part of the piston 14 forms a flow passage 2, and a non-return mechanism 13 is arranged in the flow passage 2; a plurality of layers of silicate filter paper 9 are arranged at the inner bottom of the outer cylinder 7; the bottom of the outer barrel 7 is provided with a liquid outlet 8.
The check mechanism 13 is shown in fig. 6, where the check mechanism 13 includes a support column 15 fixed in the runner, and the outer diameter of the support column 15 is smaller than the inner diameter of the runner; the spring 11 is sleeved outside the support column 15, the support column 15 is provided with a closing head 16, the closing head 16 is in sliding connection with the support column 15, and the sliding connection position of the closing head 16 and the support column 15 is located above the spring 11. Under the action of gravity of the liquid, the closing head 16 slides along the support column, and after the liquid flows down, the closing head returns to the original position under the action of the spring.
A method of extracting nucleic acid using a nucleic acid extraction consumable comprising the steps of:
1) Injecting sample solution into the outer cylinder through the liquid injection port, wherein the sample solution enters silicate filter paper;
2) Injecting the pyrolysis liquid into the outer cylinder from the liquid injection port, and allowing the pyrolysis liquid to enter the silicate filter paper
3) After the cracking is finished, the nucleic acid molecules are adsorbed by the silicon dioxide groups on the silicate filter paper, the piston is moved downwards, the piston is continuously extruded downwards after contacting the silicate filter paper, the waste liquid of the cracking liquid is discharged from the liquid outlet at the bottom, and the piston returns to the initial position after the waste liquid of the cracking liquid is discharged completely; the non-return mechanism prevents the solution from flowing back in the process of returning the piston to the initial position;
4) Injecting rinsing liquid into the outer cylinder through the liquid injection port, wherein the rinsing liquid enters silicate filter paper;
5) After standing for a period of time, moving the piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the waste liquid of the rinsing liquid at a liquid outlet at the bottom, and returning the piston to the initial position after the waste liquid of the rinsing liquid is discharged completely; the non-return mechanism prevents the solution from flowing back in the process of returning the piston to the initial position;
6) Repeating step 4) and step 5) for several times;
7) Injecting eluent into the outer cylinder from the liquid injection port, and allowing the eluent to enter silicate filter paper;
8) Standing for a period of time, eluting the nucleic acid molecules adsorbed on the silicate filter paper, moving the piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the eluent and the eluted nucleic acid molecules at the bottom liquid outlet, and dripping into a sample collector; after the eluent is discharged, the piston returns to the initial position; the non-return mechanism prevents back flow of the solution during return of the piston to the initial position.
In other embodiments, the check mechanism may be other mechanisms that prevent backflow of liquid. The matching mode of the piston and the outer cylinder is similar to that of the existing injection syringe and the syringe piston.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention, and are intended to be included within the scope of the appended claims and description.
Claims (7)
1. A nucleic acid extraction consumable, characterized in that: the device comprises an outer cylinder and a piston, wherein the piston is matched with the outer cylinder and can move up and down along the outer cylinder; the piston is hollow, and the upper opening of the hollow part of the piston is a liquid injection opening; the hollow part of the piston forms a flow passage, and a non-return mechanism is arranged in the flow passage; a plurality of layers of silicate filter paper are arranged at the inner bottom of the outer cylinder; a liquid outlet is formed in the bottom of the outer cylinder;
a sealing ring is arranged between the piston and the inner wall of the outer cylinder;
The upper part of the outer cylinder is provided with an opening, and the piston comprises an upper piston and a lower piston; the upper piston can slide up and down along the upper opening of the outer cylinder; the lower piston can slide up and down along the inner wall of the outer cylinder and is in sealing connection with the inner wall of the outer cylinder; the upper piston and the lower piston are connected into a whole; the upper piston and the lower piston are hollow;
the runner at least comprises a first sub-runner and a second sub-runner from top to bottom, and the inner diameter of the second sub-runner is larger than that of the first sub-runner.
2. The nucleic acid extraction consumable of claim 1, wherein: the check mechanism comprises a spring and a sealing ball fixed on the spring, and the bottom of the spring is fixed in the flow passage; the original position of the sealing ball is the lowest part of the first sub-runner with the top sealed.
3. The nucleic acid extraction consumable of claim 2, wherein: the flow channel consists of a first flow dividing channel, a second flow dividing channel, a third flow dividing channel and a fourth flow dividing channel which are distributed from top to bottom; the inner diameter of the third sub-runner is smaller than that of the second sub-runner; the inner diameter of the fourth sub-runner is smaller than that of the third sub-runner.
4. The nucleic acid extraction consumable of claim 1, wherein: a first sealing ring is arranged between the upper piston and the lower piston; and a second sealing ring is arranged between the lower piston and the inner wall of the outer cylinder.
5. The nucleic acid extraction consumable of claim 1, wherein: the check mechanism comprises a support column fixed in the runner, a spring is sleeved outside the support column, and a sealing head is connected to the support column in a sliding manner; the sliding connection part of the closing head and the supporting column is positioned above the spring.
6. A method for extracting nucleic acid using the nucleic acid extraction consumable according to claim 1, comprising the steps of:
1) Injecting sample solution into the outer cylinder through the liquid injection port, wherein the sample solution enters silicate filter paper;
2) Injecting a cracking liquid into the outer cylinder from the liquid injection port, wherein the cracking liquid enters silicate filter paper;
3) After the cracking is finished, the nucleic acid molecules are adsorbed by the silicon dioxide groups on the silicate filter paper, the piston is moved downwards, the piston is continuously extruded downwards after contacting the silicate filter paper, the waste liquid of the cracking liquid is discharged from the liquid outlet at the bottom, and the piston returns to the initial position after the waste liquid of the cracking liquid is discharged completely; the non-return mechanism prevents the solution from flowing back in the process of returning the piston to the initial position;
4) Injecting rinsing liquid into the outer cylinder through the liquid injection port, wherein the rinsing liquid enters silicate filter paper;
5) After standing for a period of time, moving the piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the waste liquid of the rinsing liquid at a liquid outlet at the bottom, and returning the piston to the initial position after the waste liquid of the rinsing liquid is discharged completely; the non-return mechanism prevents the solution from flowing back in the process of returning the piston to the initial position;
6) Repeating step 4) and step 5) for several times;
7) Injecting eluent into the outer cylinder from the liquid injection port, and allowing the eluent to enter silicate filter paper;
8) Standing for a period of time, eluting the nucleic acid molecules adsorbed on the silicate filter paper, moving the piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the eluent and the eluted nucleic acid molecules at the bottom liquid outlet, and dripping into a sample collector; after the eluent is discharged, the piston returns to the initial position; the non-return mechanism prevents back flow of the solution during return of the piston to the initial position.
7. A method of extracting nucleic acid using the nucleic acid extraction consumable of claim 2, comprising the steps of:
1) Injecting sample solution into the outer cylinder through the liquid injection port, pressing the spring under the action of gravity of the sample solution, allowing the sealing ball to move downwards, allowing the sample solution to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then entering silicate filter paper, allowing the spring to rebound after the sample solution passes through the silicate filter paper, and resetting the sealing ball;
2) Injecting a pyrolysis liquid into the outer cylinder from the liquid injection port, pressing the spring under the action of gravity of the pyrolysis liquid, allowing the sealing ball to move downwards, allowing the pyrolysis liquid to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then allowing the pyrolysis liquid to enter silicate filter paper, allowing the spring to rebound after the pyrolysis liquid passes through, and allowing the sealing ball to reset;
3) After the cracking is finished, the nucleic acid molecules are adsorbed by the silicon dioxide groups on the silicate filter paper, the upper piston and the lower piston are moved downwards, the lower piston is continuously extruded downwards after the nucleic acid molecules contact the silicate filter paper, the waste liquid of the cracking liquid is discharged from the liquid outlet at the bottom, and after the waste liquid of the cracking liquid is discharged, the upper piston and the lower piston are moved upwards to return to the initial position;
4) Injecting rinsing liquid into the outer cylinder from the liquid injection port, pressing the spring under the action of the gravity of the rinsing liquid, allowing the sealing ball to move downwards, allowing the rinsing liquid to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then allowing the rinsing liquid to enter silicate filter paper, allowing the spring to rebound after the rinsing liquid passes through the sealing ball, and allowing the sealing ball to reset;
5) After standing for a period of time, moving the upper piston and the lower piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the waste liquid of the rinsing liquid at a liquid outlet at the bottom, and moving the upper piston and the lower piston upwards to return to the initial position after the waste liquid of the rinsing liquid is discharged completely;
6) Repeating step 4) and step 5) for several times;
7) Injecting eluent into the outer cylinder from the liquid injection port, under the action of gravity of the eluent, pressing the spring, moving the sealing ball downwards, enabling the eluent to flow into the liquid storage area through a gap between the sealing ball and the upper piston, then entering silicate filter paper, and returning the sealing ball after the eluent passes through the sealing ball;
8) Standing for a period of time, after the nucleic acid molecules adsorbed on the silicate filter paper are eluted, moving the upper piston and the lower piston downwards, continuously extruding downwards after contacting the silicate filter paper, discharging the eluent and the eluted nucleic acid molecules at a liquid outlet at the bottom, and dripping the eluent and the eluted nucleic acid molecules into a sample collector; after the eluent is discharged, the upper piston and the lower piston are moved upwards to return to the initial positions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010668889.6A CN113930318B (en) | 2020-07-13 | 2020-07-13 | Nucleic acid extraction consumables and method for extracting nucleic acid using the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010668889.6A CN113930318B (en) | 2020-07-13 | 2020-07-13 | Nucleic acid extraction consumables and method for extracting nucleic acid using the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113930318A CN113930318A (en) | 2022-01-14 |
| CN113930318B true CN113930318B (en) | 2024-11-12 |
Family
ID=79273363
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010668889.6A Active CN113930318B (en) | 2020-07-13 | 2020-07-13 | Nucleic acid extraction consumables and method for extracting nucleic acid using the same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113930318B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN212222970U (en) * | 2020-07-13 | 2020-12-25 | 山东凯景生物技术有限公司 | Nucleic acid extraction consumptive material |
Family Cites Families (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR200217928Y1 (en) * | 2000-06-24 | 2001-03-15 | 조휘철 | pumping equipment of a container |
| KR20030018348A (en) * | 2001-08-28 | 2003-03-06 | 문인환 | A liquid fixed quantity pour into device |
| JP2005164336A (en) * | 2003-12-01 | 2005-06-23 | Aloka Co Ltd | Syringe |
| CN2720218Y (en) * | 2004-08-09 | 2005-08-24 | 朱友清 | Improved structure of one-way valve of spraying machine |
| DE102005003563B4 (en) * | 2005-01-25 | 2012-01-26 | Klebchemie, M.G. Becker Gmbh & Co Kg | metering press |
| CN203947102U (en) * | 2013-07-12 | 2014-11-19 | 中国人民解放军疾病预防控制所 | A kind of device for nucleic acid rapid extraction |
| CN105521650A (en) * | 2014-09-30 | 2016-04-27 | 宝钢不锈钢有限公司 | Quantitative gas-liquid mixing device and method |
| US9145581B1 (en) * | 2014-10-17 | 2015-09-29 | Daniel Lai | Rapid nucleic acid extraction method and apparatus |
| JP2016116469A (en) * | 2014-12-19 | 2016-06-30 | セイコーエプソン株式会社 | Cartridge for purifying organism-related substance |
| US9957500B2 (en) * | 2015-02-26 | 2018-05-01 | Stem Arts Projects, Llc | Continuous flow nucleic acid extraction apparatus and method of use thereof |
| CN105296327B (en) * | 2015-10-21 | 2017-10-27 | 陈辉 | A kind of nucleic acid-extracting apparatus and its extracting method |
| CN105214343B (en) * | 2015-10-26 | 2017-06-20 | 张家港高品诚医械科技有限公司 | A kind of extraction suit of platelet rich plasma |
| CN206705760U (en) * | 2016-10-17 | 2017-12-05 | 成都川力智能流体设备股份有限公司 | A kind of pipeline Antimicrobial device |
| CN206308350U (en) * | 2016-11-22 | 2017-07-07 | 李艳芬 | A kind of biogas is produced with the water circulation cleaning device of raw material stalk |
| CN106701548A (en) * | 2017-02-10 | 2017-05-24 | 柳州市妇幼保健院 | Lymphocyte extraction device and extraction method |
-
2020
- 2020-07-13 CN CN202010668889.6A patent/CN113930318B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN212222970U (en) * | 2020-07-13 | 2020-12-25 | 山东凯景生物技术有限公司 | Nucleic acid extraction consumptive material |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113930318A (en) | 2022-01-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN212222970U (en) | Nucleic acid extraction consumptive material | |
| US9927342B2 (en) | Two station sample and washing system | |
| CN104807826A (en) | Self-cleaning automatic water sample inlet device | |
| CN113930318B (en) | Nucleic acid extraction consumables and method for extracting nucleic acid using the same | |
| CN201431985Y (en) | Cleaning device used for cleaning outer wall of sampling needle | |
| CN105733935A (en) | Preparation and detection reagent card case for nucleic acid or protein | |
| CN110479392A (en) | A kind of microfluidic valve and its detection chip | |
| CN118150746B (en) | Gas chromatograph sample leading-in device | |
| CN202204701U (en) | Quantitative capillary tube | |
| CN107063771B (en) | Dialysis machine inlet water sampler | |
| TWM638918U (en) | Multi-pass valve assembly | |
| CN219897904U (en) | Mixed heating device | |
| CN216170104U (en) | Extraction device for chemical experiment | |
| CN110732158A (en) | supercritical CO of wall-broken Ganoderma spore powder2Fluid extraction equipment | |
| CN115895833A (en) | Kit based on gravity-driven liquid flow and detection method thereof | |
| CN114990361A (en) | Multistage filter-pressing membrane-breaking mercury purification device and method under normal temperature condition | |
| CN205413065U (en) | A device for separating solution component | |
| CN110025884A (en) | A kind of female Luer | |
| CN206622129U (en) | It is a kind of can liquid separation centrifuge tube | |
| CN208466744U (en) | Sample introduction needle cleaning device | |
| CN206232701U (en) | Magnetic frame with liquid suction function | |
| CN110261636B (en) | Liquid sampling device | |
| CN208927656U (en) | A glass column chromatography sample loading device | |
| CN112891986B (en) | Integrated closed nucleic acid automatic extraction device | |
| CN110437980B (en) | Nucleic acid extraction adsorption column |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |