CN112961903A - Inactivated virus sample nucleic acid preservation solution and preparation method thereof - Google Patents
Inactivated virus sample nucleic acid preservation solution and preparation method thereof Download PDFInfo
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- CN112961903A CN112961903A CN202110333199.XA CN202110333199A CN112961903A CN 112961903 A CN112961903 A CN 112961903A CN 202110333199 A CN202110333199 A CN 202110333199A CN 112961903 A CN112961903 A CN 112961903A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F27/00—Mixers with rotary stirring devices in fixed receptacles; Kneaders
- B01F27/80—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis
- B01F27/90—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis with paddles or arms
- B01F27/906—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis with paddles or arms with fixed axis
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/20—Measuring; Control or regulation
- B01F35/22—Control or regulation
- B01F35/221—Control or regulation of operational parameters, e.g. level of material in the mixer, temperature or pressure
- B01F35/2215—Temperature
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Abstract
The invention discloses an inactivated virus sample nucleic acid preservation solution and a preparation method thereof, and relates to the technical field of biological preservation solution preparation, wherein the nucleic acid preservation solution comprises the following components in percentage by weight: 13-35% of lithium dodecyl sulfate, 35-50% of N-lauroyl sarcosine, 5-10% of sodium citrate, 25-40% of buffer solution and the balance of double distilled water, wherein the method comprises the following steps: s100: proportioning the components of the nucleic acid preservation solution according to the weight percentage; s200: placing the components proportioned in the step S100 in a mixing mechanism for mixing; s300: taking out the mixed solution in the step S200 to prepare a nucleic acid preservation solution; the mixing mechanism comprises a storage system for placing nucleic acid preservation solution components, a driving system arranged on the surface of the storage system and a stirring assembly arranged in the storage system, and the driving system drives the stirring assembly to rotate.
Description
Technical Field
The invention relates to the technical field of preparation of biological preservation solution, in particular to inactivated virus sample nucleic acid preservation solution and a preparation method thereof.
Background
Viruses are special organisms without cellular structures, they are very simple in structure, consist of protein coat and internal genetic material, DNA or RNA, and are extremely minute in size and need to be replicated in living host cells. The virus can infect human, animal, plant, bacteria and fungi, can cause host diseases, especially respiratory tract infectious viruses such as SARS, A type H1N1 and coronavirus, and has strong infectivity, thereby causing great harm to human health.
In order to determine whether the virus is infected by the virus, a sample is generally required to be sampled for analysis and determination, the activity of the virus is kept in a short time after sampling, the sample swab can be used for molecular biological detection of the virus and separation and culture of the virus, after the virus sample is collected, in order to maintain the activity of the virus sample and prolong the survival time of the virus in the sample, the sample swab is put into a preservation solution for preservation and transportation, because human operation is required during detection and experiment of the virus, if the virus is in an active state at the moment, the infection of an operator is easy, and for the reasons, some virus samples are firstly inactivated before transportation and then put into the preservation solution for preservation, but no preservation solution in the prior art can preserve the inactivated virus sample.
Therefore, compared with the prior art, the method can prepare the preservation solution more efficiently, and the preservation solution can better preserve the virus sample.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides an inactivated virus sample nucleic acid preservation solution and a preparation method thereof.
The invention is realized by the following technical scheme: the nucleic acid preservation solution for the inactivated virus sample comprises the following components in percentage by weight: 13-35% of lithium dodecyl sulfate, 35-50% of N-lauroyl sarcosine, 5-10% of sodium citrate, 25-40% of buffer solution and the balance of double distilled water.
Preferably, the buffer solution consists of the following components in percentage by weight: 25 to 36 percent of sodium chloride, 30 to 35 percent of magnesium chloride, 13 to 21 percent of sodium citrate and the balance of DEPC water.
Preferably, the preservation solution consists of the following components in percentage by weight: 18% of lithium dodecyl sulfate, 35% of N-lauroyl sarcosine, 9% of sodium citrate, 22% of buffer solution and 16% of double distilled water.
Preferably, the buffer solution consists of the following components in percentage by weight: 30% sodium chloride, 30% magnesium chloride, 18% sodium citrate and 22% DEPC water.
The invention also discloses a preparation method of the nucleic acid preservation solution for the inactivated virus sample, which comprises the following steps:
s100: proportioning the components of the nucleic acid preservation solution according to the weight percentage;
s200: placing the components proportioned in the step S100 in a mixing mechanism for mixing;
s300: taking out the solution mixed in the step S200 to prepare the nucleic acid preservation solution;
wherein, the mixing mechanism comprises a storage system for placing nucleic acid preservation solution components, a driving system arranged on the surface of the storage system and a stirring assembly arranged in the storage system, and the driving system drives the stirring assembly to rotate.
Based on the above process, for the preparation process and the device of the invention, the mixing mechanism is used for mixing different proportioning components, the driving mechanism is used for driving the stirring component to rotate, and the components with different proportioning components can be uniformly mixed together through the rotation of the stirring component.
Preferably, the storage system comprises a storage tank and a sleeve body provided with a storage groove, and the inner wall of the storage groove of the sleeve body is in contact with the outer wall of the storage tank; the sleeve body is arranged into a hollow structure, and a liquid inlet and a liquid outlet are formed in the surface of the sleeve body;
the storage system has a temperature adjusting function, liquid is supplied to the sleeve body through liquid inlet by external liquid supply equipment, the temperature of liquid in the sleeve body is transferred to the storage tank in a heat transfer mode, and the temperature of the liquid stored in the storage tank is adjusted.
Preferably, to enhance the effect of mixing, the stirring assembly comprises a shaft arranged to be driven in rotation by the drive mechanism; at least one stirring blade arranged to be connected to the rotating shaft; the stirring blades can move in the axial direction of the rotating shaft, the heights of the stirring blades can be adjusted according to the height of liquid in the storage tank, all the stirring blades are submerged in the liquid, the area of the stirring blades is reduced, the running resistance of the stirring blades in the rotating process is reduced, the use of raw materials is saved, and the mixing efficiency of the mixing mechanism is improved.
Preferably, in order to facilitate understanding, the invention also discloses a specific structure for connecting the rotating shaft and the stirring blade, wherein a groove body extending along the axis direction is formed in the circumferential direction of the surface of the rotating shaft; the root of the stirring blade is provided with a limiting block, the limiting block is connected with the groove body in a sliding mode, and the surface of the limiting block is provided with a threaded through hole along the direction parallel to the axis of the rotating shaft; wherein, threaded rod threaded connection the screw thread through-hole, just threaded rod one end is rotated and is connected the cell body bottom.
Preferably, in order to make the stirring blade more stable in the operation process, one end of the stirring blade is movably connected with the stabilizing member through a rotating shaft, and the stabilizing member comprises: a first support arm; a second support arm arranged to connect the first support arm; the roller is arranged to be rotatably arranged on one side, close to the inner wall of the storage tank, of the second supporting arm; wherein, the material of storage jar is magnetic metal, the material of second support arm is magnetic adsorption mechanism.
The invention discloses an inactivated virus sample nucleic acid preservation solution and a preparation method thereof, compared with the prior art:
the method can prepare the preservation solution more efficiently, the preservation solution can also better preserve the virus sample, for the preparation process and the preparation device, components with different proportions are mixed through the mixing mechanism, for the mixing mechanism, the storage system is used for containing the components of the nucleic acid cell sap, the driving mechanism drives the stirring assembly to rotate, the driving mechanism can be arranged at the bottom of the storage system, the structural part of the driving mechanism comprises a motor and a gearbox connected with the driving motor, the components with different proportions can be uniformly mixed together through the rotation of the stirring assembly, the supporting system comprises supporting legs and universal wheels arranged at the bottoms of the supporting legs, and the mixing mechanism can be moved easily through the arrangement of the universal wheels.
Drawings
FIG. 1 is a front view of a mixing mechanism of the present invention;
FIG. 2 is a schematic external view of the mixing mechanism of the present invention;
FIG. 3 is a schematic structural diagram of a memory system according to the present invention;
FIG. 4 is a schematic view of the connection between the rotating shaft and the stirring blade according to the present invention;
FIG. 5 is a schematic view of the stirring assembly of the present invention;
fig. 6 is a partially enlarged schematic view of a portion a of fig. 5.
Detailed Description
The following examples are given for the detailed implementation and specific operation of the present invention, but the scope of the present invention is not limited to the following examples.
The nucleic acid preservation solution for the inactivated virus sample comprises the following components in percentage by weight: 13-35% of lithium dodecyl sulfate, 35-50% of N-lauroyl sarcosine, 5-10% of sodium citrate, 25-40% of buffer solution and the balance of double distilled water.
The buffer solution comprises the following components in percentage by weight: 25 to 36 percent of sodium chloride, 30 to 35 percent of magnesium chloride, 13 to 21 percent of sodium citrate and the balance of DEPC water.
For ease of understanding, specific examples of buffer and preservative solutions are disclosed.
The nucleic acid preservation solution comprises the following components in percentage by weight: 18% of lithium dodecyl sulfate, 35% of N-lauroyl sarcosine, 9% of sodium citrate, 22% of buffer solution and 16% of double distilled water; the buffer solution comprises the following components in percentage by weight: 30% sodium chloride, 30% magnesium chloride, 18% sodium citrate and 22% DEPC water.
The invention also discloses a preparation method of the nucleic acid preservation solution for the inactivated virus sample, which comprises the following steps:
s100: proportioning the components of the nucleic acid preservation solution according to the weight percentage;
s200: placing the components proportioned in the step S100 in a mixing mechanism for mixing;
s300: taking out the solution mixed in the step S200 to prepare the nucleic acid preservation solution;
as shown in fig. 1 and 2, the mixing mechanism includes a storage system 1 for storing the components of the nucleic acid preservation solution, a driving system 2 disposed on the surface of the storage system 1, and a stirring assembly 3 disposed inside the storage system 1, wherein the driving system 2 drives the stirring assembly 3 to rotate, and a support system 4 is further disposed at the bottom of the storage system 1 to support the storage system 1.
Based on the above process, for the preparation process and apparatus of the present invention, different mixture ratio components are mixed by a mixing mechanism, for the mixing mechanism, a storage system 1 is used for containing components of nucleic acid cell liquid, a driving mechanism 2 drives a stirring component 3 to rotate, the driving mechanism 2 can be arranged at the bottom of the storage system 1, the structural component of the mixing mechanism comprises a motor 2 and a gearbox connected with the driving motor, different mixture ratio components can be uniformly mixed together by the rotation of the stirring component 3, the supporting system 4 comprises supporting legs and universal wheels arranged at the bottoms of the supporting legs, and the mixing mechanism of the present invention can be easily moved by the arrangement of the universal wheels.
In order to adjust the temperature of the liquid during mixing by the storage system 1 during the operation of the mixing mechanism, as shown in fig. 3, the storage system 1 includes a storage tank 11 and a cover body 12 provided with a storage groove, and the inner wall of the storage groove of the cover body 12 contacts the outer wall of the storage tank 11; wherein the sleeve body 12 is arranged in a hollow structure, and a liquid inlet and a liquid outlet are arranged on the surface of the sleeve body;
as for the storage system 1, it has a temperature adjusting function, an external liquid supply device supplies liquid to the sleeve 12 through liquid inlet, the liquid in the sleeve 12 transfers the temperature to the storage tank 11 through heat transfer, and the temperature of the liquid stored in the storage tank 11 is adjusted, wherein, as shown in fig. 2, the external liquid supply device may be connected with a liquid feeding pipe 5 of a hot water liquid supply system, the liquid feeding pipe 5 is in a communication state with the storage tank 11, so that the heated liquid can enter the cavity of the sleeve 12 through the liquid feeding pipe 5, wherein, the sleeve 12 adopts a layered design, the cavity is internally divided by a partition plate from top to bottom to form an independent space, one independent space corresponds to one liquid feeding pipe, and the dissipation of the liquid temperature in a large space can be reduced through the arrangement of the independent space.
In order to enhance the mixing effect, the stirring assembly 3 comprises a rotating shaft 31 arranged to be driven in rotation by the driving mechanism 2; at least one stirring blade 32 arranged to be connected to the rotation shaft 31; the stirring blades 32 can move along the axial direction of the rotating shaft 32, and the height of the stirring blades 32 can be adjusted according to the height of liquid in the storage tank 11, so that all the stirring blades are submerged in the liquid, the area of the stirring blades 32 is reduced, the air resistance of the stirring blades 32 in the rotating process is reduced, the use of raw materials is saved, and the mixing efficiency of the mixing mechanism is improved.
For convenience of understanding, the present invention further discloses a specific embodiment of the connection between the rotating shaft 31 and the stirring blade 32, as shown in fig. 4, a groove 311 extending along the axial direction is formed on the circumferential direction of the surface of the rotating shaft 31; a limiting block 321 is arranged at the root of the stirring blade 32, the limiting block 321 is connected with the groove body 311 in a sliding manner, and a threaded through hole is formed in the surface of the limiting block 321 along a direction parallel to the axis of the rotating shaft 31; wherein, threaded rod threaded connection the screw thread through-hole, just threaded rod one end is rotated and is connected the cell body 311 bottom rotates the threaded rod, and stopper 321 removes in cell body 311 is inside to drive the removal of stirring leaf 32, accomplish the regulation to stirring leaf position, stopper 321 can set to circular-arcly, thereby increase and the cell body 311 between area of contact, more stable.
Because the limiting block 321 drives the stirring blade 32 to slide in the slot 311, the limiting block 321 may shake during the rotation of the rotating shaft 31, in order to make the stirring blade 32 more stable during the operation, as shown in fig. 5 and 6, one end of the stirring blade 32 is movably connected to the stabilizing member 33 through the rotating shaft, and the stabilizing member 33 includes: the first support arm 331; a second support arm 332 arranged to connect the first support arm 331; a roller 333 arranged to be rotatably provided on a side of the second support arm 332 close to the inner wall of the storage tank 11; wherein, the material of storage jar 11 is magnetic metal, the material of second support arm 332 is magnetic adsorption mechanism, and magnetic mechanism can be electro-magnet or magnetite, and sliding connection between first support arm 331 and the second support arm, and then can make stabilizer 33 can be applicable to not unidimensional storage system 1.
As described above, the method of the present invention allows the preparation of a storage solution more efficiently than the conventional methods, and the storage solution can also store a virus sample more favorably, and it is noted that the preparation method of the present invention can be used for the preparation of not only the nucleic acid storage solution of the present invention but also other nucleic acid storage solutions which need to be mixed well.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention and the equivalent alternatives or modifications according to the technical solution and the inventive concept of the present invention within the technical scope of the present invention.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
Claims (10)
1. The nucleic acid preservation solution for the inactivated virus sample is characterized by comprising the following components in percentage by weight: 13-35% of lithium dodecyl sulfate, 35-50% of N-lauroyl sarcosine, 5-10% of sodium citrate, 25-40% of buffer solution and the balance of double distilled water.
2. The preservation solution for nucleic acid in inactivated virus samples according to claim 1, wherein the buffer solution comprises the following components by weight percent: 25 to 36 percent of sodium chloride, 30 to 35 percent of magnesium chloride, 13 to 21 percent of sodium citrate and the balance of DEPC water.
3. The preservation solution for nucleic acid of inactivated virus sample according to claim 1, which comprises the following components by weight percent: 18% of lithium dodecyl sulfate, 35% of N-lauroyl sarcosine, 9% of sodium citrate, 22% of buffer solution and 16% of double distilled water.
4. The preservation solution for nucleic acid in inactivated virus samples according to claim 2, wherein the buffer solution comprises the following components by weight percent: 30% sodium chloride, 30% magnesium chloride, 18% sodium citrate and 22% DEPC water.
5. A method for preparing a nucleic acid preservation solution for an inactivated virus sample, which comprises preparing the nucleic acid preservation solution according to any one of claims 1 to 4.
6. The method for preparing a nucleic acid preservation solution for inactivated virus samples according to claim 5, wherein the method comprises the steps of:
s100: proportioning the components of the nucleic acid preservation solution according to the weight percentage;
s200: placing the components proportioned in the step S100 in a mixing mechanism for mixing;
s300: taking out the solution mixed in the step S200 to prepare the nucleic acid preservation solution;
wherein, the mixing mechanism comprises a storage system (1) for placing the components of the nucleic acid preservation solution, a driving system (2) arranged on the surface of the storage system (1) and a stirring component (3) arranged inside the storage system (1), and the driving system (2) drives the stirring component (3) to rotate.
7. The method for preparing the nucleic acid preservation solution for inactivated virus samples according to claim 6, wherein the storage system (1) comprises a storage tank (11) and a sleeve body (12) provided with a storage groove, and an inner wall of the storage groove of the sleeve body (12) contacts with an outer wall of the storage tank (11);
wherein, the sleeve body (12) is arranged into a hollow structure, and the surface of the sleeve body is provided with a liquid inlet and a liquid outlet.
8. The method for preparing a nucleic acid preservation solution for inactivated virus samples according to claim 7, wherein the stirring module (3) comprises:
a rotating shaft (31) arranged to be driven in rotation by the drive mechanism (2);
at least one stirring blade (32) arranged in connection with the shaft (31);
wherein, the stirring blade (32) can move along the axial direction of the rotating shaft (32).
9. The method for preparing the nucleic acid preservation solution for the inactivated virus sample according to claim 8, wherein the surface of the rotating shaft (31) is circumferentially provided with a groove (311) extending along the axial direction;
the root of the stirring blade (32) is provided with a limiting block (321), the limiting block (321) is connected with the groove body (311) in a sliding manner, and the surface of the limiting block (321) is provided with a threaded through hole along the direction parallel to the axis of the rotating shaft (31);
wherein, threaded rod threaded connection the screw thread through-hole, just threaded rod one end is rotated and is connected bottom slot body (311).
10. The method for preparing a nucleic acid-preserving fluid for inactivated virus samples according to claim 9, wherein one end of the stirring blade (32) is movably connected to the stabilizer (33) through a rotating shaft, and the stabilizer (33) comprises:
a first support arm (331);
a second support arm (332) arranged to connect the first support arm (331);
a roller (333) arranged to be rotatably provided on a side of the second support arm (332) close to an inner wall of the storage tank (11);
wherein, the material of storage jar (11) is magnetic metal, the material of second support arm (332) is magnetic adsorption mechanism.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110333199.XA CN112961903B (en) | 2021-03-29 | 2021-03-29 | Inactivated virus sample nucleic acid preservation solution and preparation method thereof |
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| CN202110333199.XA CN112961903B (en) | 2021-03-29 | 2021-03-29 | Inactivated virus sample nucleic acid preservation solution and preparation method thereof |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103506033A (en) * | 2012-06-21 | 2014-01-15 | 四川制药制剂有限公司 | Efficient stirring system used for pharmaceutical workshop |
| CN206500078U (en) * | 2017-02-27 | 2017-09-19 | 四川德成动物保健品有限公司 | Drug solution preparing tank based on removable agitating paddle |
| CN111676216A (en) * | 2020-06-18 | 2020-09-18 | 合肥铼科生物科技有限公司 | Extraction-free inactivated virus sample nucleic acid preservation solution and preparation method and application thereof |
| CN111718908A (en) * | 2020-08-11 | 2020-09-29 | 杭州博日科技有限公司 | Virus sample preserving fluid and preparation method and application thereof |
| US20210071232A1 (en) * | 2018-11-14 | 2021-03-11 | Spectrum Solutions L.L.C. | Rna preservation solution and methods of manufacture and use |
-
2021
- 2021-03-29 CN CN202110333199.XA patent/CN112961903B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103506033A (en) * | 2012-06-21 | 2014-01-15 | 四川制药制剂有限公司 | Efficient stirring system used for pharmaceutical workshop |
| CN206500078U (en) * | 2017-02-27 | 2017-09-19 | 四川德成动物保健品有限公司 | Drug solution preparing tank based on removable agitating paddle |
| US20210071232A1 (en) * | 2018-11-14 | 2021-03-11 | Spectrum Solutions L.L.C. | Rna preservation solution and methods of manufacture and use |
| CN111676216A (en) * | 2020-06-18 | 2020-09-18 | 合肥铼科生物科技有限公司 | Extraction-free inactivated virus sample nucleic acid preservation solution and preparation method and application thereof |
| CN111718908A (en) * | 2020-08-11 | 2020-09-29 | 杭州博日科技有限公司 | Virus sample preserving fluid and preparation method and application thereof |
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| CN112961903B (en) | 2022-11-01 |
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