CN112725413B - Splitting and combining liquid, kit and method for extracting nucleic acid from cervical swab - Google Patents
Splitting and combining liquid, kit and method for extracting nucleic acid from cervical swab Download PDFInfo
- Publication number
- CN112725413B CN112725413B CN202110163794.3A CN202110163794A CN112725413B CN 112725413 B CN112725413 B CN 112725413B CN 202110163794 A CN202110163794 A CN 202110163794A CN 112725413 B CN112725413 B CN 112725413B
- Authority
- CN
- China
- Prior art keywords
- centrifuge tube
- liquid
- magnetic
- nucleic acid
- tube
- 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
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of nucleic acid extraction, and discloses a lysis binding solution, a kit and a method for extracting nucleic acid from a cervical swab, wherein the lysis binding solution comprises the following components in concentration: 1-5% of sodium dodecyl sulfate, 0.5-2% of polyethylene glycol octyl phenyl ether, 100-300mM of 4-hydroxyethyl piperazine ethanesulfonic acid, 8-12% of sodium chloride, 50-150mM of tris (hydroxymethyl) aminomethane hydrochloride, 0.5-3mM of EDTA and 1-3% of sodium lauryl polyoxyethylene ether sulfate.
Description
Technical Field
The invention relates to the technical field of nucleic acid extraction, in particular to a lysis binding solution, a kit and a method for extracting nucleic acid from a cervical swab.
Background
With the development of medical technology, there is an increasing concern about female health. Before the diagnosis and treatment of gynecological diseases, cervical cells and body fluid are collected by using a cervical swab, and then the collected cells and body fluid are analyzed to obtain medical judgment and make a treatment scheme.
The existing nucleic acid extraction methods mainly comprise a pyrolysis method, a phenol chloroform extraction method, a centrifugal column method and a magnetic bead method; the magnetic bead method adopts superparamagnetic nanoparticles, nucleic acid is adsorbed under high salt and low pH, and then the nucleic acid is separated and purified under the principles of low salt and high pH, and the steps of nucleic acid extraction by the magnetic bead method mainly comprise: (1) cracking; (2) binding; (3) washing; (4) Separating four major parts, specifically, under the action of a lysis solution, specifically, after releasing DNA/RNA, carrying out specific binding with surface-modified superparamagnetic nano magnetic beads to form a nucleic acid-magnetic bead compound, then adding a washing solution to wash away impurities such as nonspecifically adsorbed proteins, finally, separating and enriching magnetic beads under the condition of a magnetic field, and finally, eluting nucleic acid from the magnetic beads by using an eluent to obtain the DNA/RNA to be extracted. The magnetic bead method can separate and purify nucleic acid only under the action of a magnetic field, so that high-throughput automatic standardized operation can be realized.
However, the commercial reagents based on the magnetic bead method in the current market are too complex to operate, require incubation with proteinase K and the like at a certain temperature, and require multiple washing steps, which results in low automation degree, poor nucleic acid extraction repeatability and low extraction efficiency.
Disclosure of Invention
Therefore, a lysis binding solution, a kit and a method for extracting nucleic acid from a cervical swab are needed to be provided, so that the problems of complicated steps, low extraction rate and the like in the existing nucleic acid extraction process are solved.
In order to achieve the above object, the present invention provides a lysis binding solution for cervical swab nucleic acid extraction, comprising the following components in concentration: 1-5% of sodium dodecyl sulfate, 0.5-2% of polyethylene glycol octyl phenyl ether, 100-300mM of 4-hydroxyethyl piperazine ethanesulfonic acid, 8-12% of sodium chloride, 50-150mM of tris (hydroxymethyl) aminomethane hydrochloride, 0.5-3mM of EDTA and 1-3% of sodium lauryl polyoxyethylene ether sulfate.
Further, the following components were included in the following concentrations: 2% of sodium dodecyl sulfate, 1% of polyethylene glycol octyl phenyl ether, 200mM 4-hydroxyethyl piperazine ethanesulfonic acid, 10% of sodium chloride, 100mM of tris (hydroxymethyl) aminomethane hydrochloride, 1mM of EDTA and 2% of sodium lauryl polyoxyethylene ether sulfate.
Further, the pH value of the cracking combination liquid is kept between 7.0 and 8.0.
A cervical swab nucleic acid extraction kit comprises the lysis binding solution and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads and a preservative, the magnetic beads are used for adsorbing DNA, and the concentration of the magnetic bead solution is 200-400 mu g/ml.
Further, the amount of the magnetic beads was 300. Mu.g/ml.
A method of cervical swab nucleic acid extraction, comprising the steps of:
(1) Adding the lysis binding solution and the magnetic bead solution into a centrifuge tube, uniformly mixing, then adding a cervical swab sample, uniformly mixing by vortex oscillation for 5-20s, and then standing for 1-2min in a water bath at 70-95 ℃, wherein the volume ratio of the magnetic bead solution to the lysis binding solution is 1-20-40, and the volume ratio of the cervical swab sample to the lysis binding solution is 1-2-5;
(2) Taking out the centrifugal tube in the step (1) from the water bath, standing, and cooling to below 25 ℃;
(3) Swirling the centrifugal tube cooled in the step (2) for 5-15s, then placing the centrifugal tube on a magnetic frame until all magnetic beads are adsorbed to the wall of the centrifugal tube and the liquid in the centrifugal tube is clear, and then absorbing and removing the liquid;
(4) Removing the magnetic frame, adding DNA eluent into the centrifuge tube, wherein the volume ratio of the NA eluent to the cervical swab sample is 1-2, blowing or suspending and oscillating the magnetic beads on the tube wall by using a pipette tip until the liquid in the centrifuge tube is uniformly distributed, and collecting the elution liquid in the centrifuge tube.
Further, in the step (1), the volume ratio of the magnetic bead solution to the lysis binding solution is 1.
Further, in the step (3), after the liquid is sucked, a step of sucking and discarding the residual liquid is performed, and the step of sucking and discarding the residual liquid is one of the following modes:
(1) Standing the centrifugal tube for 0.5-1min, allowing residual liquid to fall to the bottom of the centrifugal tube along the wall of the centrifugal tube, and sucking away the residual liquid;
(2) Instantly centrifuging the centrifuge tube, placing the centrifuge tube on a magnetic rack, and sucking and discarding residual liquid;
(3) The centrifuge tube was tapped so that the residual liquid on the wall of the centrifuge tube all fell to the bottom, and then the centrifuge tube was placed on a magnetic stand and the residual liquid was discarded.
Further, in step (4), the volume ratio of the NA eluate to the cervical swab sample is 1.
Further, the DNA eluent is deionized water, DEPC water or PCR mixed reaction solution.
The technical scheme has the following beneficial effects:
1. the lysis binding solution can complete tissue cell lysis and DNA binding in one step without incubation, simplifies the operation steps, effectively saves the operation time and labor cost, and improves the nucleic acid extraction efficiency of the cervical swab sample.
2. In the invention, HEPES (4-hydroxyethyl piperazine ethanesulfonic acid) and Tris-HCl (Tris-hydroxymethyl aminomethane hydrochloride) jointly act as a buffer solution, sodium dodecyl sulfonate, polyoxyethylene octyl phenyl ether and sodium lauryl polyoxyethylene ether sulfate have a synergistic effect on the cracking of cervical tissue cells, and the concentration and quality of extracted DNA can be greatly improved by the synergistic use, so that the nucleic acid extraction efficiency of the cervical swab is improved.
Detailed Description
In order to explain technical contents, structural features, and objects and effects of the technical means in detail, the following detailed description is given with reference to specific embodiments.
Example 1
A lysis binding solution for cervical swab nucleic acid extraction comprising the following components in concentrations: 2% sodium dodecyl sulfate, 1% polyoxyethylene octyl phenyl ether, 200mM 4-hydroxyethyl piperazine ethanesulfonic acid, 10% sodium chloride, 100mM tris (hydroxymethyl) aminomethane hydrochloride, 1mM EDTA and 2% sodium lauryl polyoxyethylene ether sulfate. The pH value of the cracking combination liquid is kept between 7.0 and 8.0.
A kit for extracting nucleic acid from a cervical swab comprises the lysis binding solution and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads and a preservative, the magnetic beads are used for adsorbing DNA, and the dosage of the magnetic beads is 300 mu g/ml.
A method of cervical swab nucleic acid extraction, comprising the steps of:
(1) Adding 600 mu L of the lysis binding solution and 20 mu L of the magnetic bead solution into a micro-centrifuge tube, mixing uniformly, then adding 200 mu L of the cervical swab sample, mixing uniformly by vortex oscillation for 10s, and then standing for 1min in a water bath at 90 ℃;
(2) Taking out the centrifugal tube in the step (1) from the water bath, standing, cooling to below 25 ℃, wherein the cooling mode is natural cooling or placing the centrifugal tube in cold water at the temperature of 25 ℃ for accelerated cooling;
(3) Swirling the cooled centrifugal tube in the step (2) for 10s, then placing the centrifugal tube on a magnetic frame, standing for 4min until all magnetic beads are adsorbed to the wall of the centrifugal tube and the liquid in the centrifugal tube is clarified, absorbing the liquid, wherein the magnetic beads are not taken out when the liquid is absorbed, and in order to absorb all the liquid to the greatest extent, the residual liquid in the centrifugal tube is also absorbed, in the embodiment, the liquid is absorbed for the first time; standing the centrifugal tube for 0.5-1min, allowing residual liquid in the centrifugal tube to fall to the bottom of the centrifugal tube along the wall of the centrifugal tube, and sucking away the residual liquid;
(4) And removing the magnetic frame, adding 50 mu L of DNA eluent into the centrifuge tube, wherein the DNA eluent is deionized water, blowing the magnetic beads in the centrifuge tube by using a pipette tip until the liquid in the centrifuge tube is uniformly distributed, and collecting the elution liquid in the centrifuge tube.
Example 2
A lysis binding solution for cervical swab nucleic acid extraction comprising the following components in concentrations: 1% sodium dodecyl sulfate, 0.5% polyoxyethylene octyl phenyl ether, 100mM 4-hydroxyethyl piperazine ethanesulfonic acid, 8% sodium chloride, 50mM tris (hydroxymethyl) aminomethane hydrochloride, 0.5mM EDTA and 1% sodium laureth sulfate. The pH value of the cracking combination liquid is kept between 7.0 and 8.0.
A kit for extracting nucleic acid from a cervical swab comprises the lysis binding solution and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads and a preservative, the magnetic beads are used for adsorbing DNA, and the dosage of the magnetic beads is 200 mu g/ml.
A method of cervical swab nucleic acid extraction, comprising the steps of:
(1) Adding 400 mu L of the lysis binding solution and 20 mu L of the magnetic bead solution into a micro-centrifuge tube, uniformly mixing, then adding 200 mu L of cervical swab sample, uniformly mixing by vortex oscillation for 5s, and then standing for 2min in a water bath at 70 ℃;
(2) Taking out the centrifugal tube in the step (1) from the water bath, standing, and cooling to below 25 ℃, wherein the cooling mode is natural cooling;
(3) Swirling the cooled centrifuge tube in the step (2) for 5s, then placing the centrifuge tube on a magnetic frame, standing for 4min until all magnetic beads are adsorbed to the wall of the centrifuge tube and the liquid in the centrifuge tube is clear, absorbing the liquid, and when the liquid is absorbed, not taking out the magnetic beads, in order to absorb all the liquid as much as possible and completely clean, then absorbing the residual liquid in the centrifuge tube, wherein in the embodiment, the liquid is absorbed for the first time; instantly centrifuging the centrifuge tube, placing the centrifuge tube on a magnetic rack, and sucking and discarding residual liquid;
(4) And removing the magnetic frame, adding 50 mu L of DNA eluent into the centrifuge tube, wherein the DNA eluent is deionized water, blowing the magnetic beads in the centrifuge tube by using a pipette tip until the liquid in the centrifuge tube is uniformly distributed, and collecting the elution liquid in the centrifuge tube.
Example 3
A lysis binding solution for cervical swab nucleic acid extraction, comprising the following components in concentration: 5% sodium dodecyl sulfate, 5% polyoxyethylene octyl phenyl ether, 300mM 4-hydroxyethyl piperazine ethanesulfonic acid, 12% sodium chloride, 150mM tris (hydroxymethyl) aminomethane hydrochloride, 3mM EDTA and 2% sodium laureth sulfate. The pH value of the cracking combination liquid is kept between 7.0 and 8.0.
A kit for extracting nucleic acid from cervical swabs comprises the lysis binding solution and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads and a preservative, the magnetic beads are used for adsorbing DNA, and the dosage of the magnetic beads is 400 mu g/ml.
A method of cervical swab nucleic acid extraction, comprising the steps of:
(1) Adding 1000. Mu.L of the lysis binding solution and 25. Mu.L of the magnetic bead solution into a micro-centrifuge tube, mixing, adding 200. Mu.L of the cervical swab sample, mixing uniformly by vortex oscillation for 20s, and standing for 1min in a water bath at 95 ℃;
(2) Taking out the centrifugal tube in the step (1) from the water bath, standing, cooling to below 25 ℃, wherein the cooling mode is natural cooling or placing the centrifugal tube in cold water at the temperature of 25 ℃ for accelerated cooling;
(3) Swirling the centrifugal tube cooled in the step (2) for 15s, then placing the centrifugal tube on a magnetic frame, standing for 4min until all magnetic beads are adsorbed to the wall of the centrifugal tube and liquid in the centrifugal tube is clarified, absorbing and discarding liquid, wherein the magnetic beads are not taken out when the liquid is absorbed and discarded, and in order to completely absorb and discard the liquid as much as possible, residual liquid in the centrifugal tube is also required to be absorbed and discarded;
(4) And removing the magnetic frame, adding 50 mu L of DNA eluent which is deionized water into the centrifugal tube, blowing magnetic beads in the centrifugal tube by using a suction head of a pipette until the liquid in the centrifugal tube is uniformly distributed, and collecting the elution liquid in the centrifugal tube.
Example 4
In this example, the difference from example 1 is that DEPC water is used as an eluent in the step (4) of the extraction method.
Example 5
The present example is different from example 1 in that, in the step (4) of the extraction method, the eluent is a mixed reaction solution of PCR or PCR.
Example 6
The preparation of the lysis solution was the same as in example 1, except for the method of extracting nucleic acid from cervical swabs in the nucleic acid kit.
The kit for extracting nucleic acid from the cervical swab comprises the lysis binding solution and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads and a preservative, the magnetic beads are used for adsorbing DNA, and the dosage of the magnetic beads is 200 mu g/ml.
The method for extracting nucleic acid from cervical swabs comprises the following steps:
(1) Adding 400 mu L of the lysis binding solution and 20 mu L of the magnetic bead solution into a micro-centrifuge tube, mixing uniformly, then adding 200 mu L of the cervical swab sample, mixing uniformly by vortex oscillation for 5s, and then standing for 2min in a water bath at 70 ℃;
(2) Taking out the centrifugal tube in the step (1) from the water bath, standing, and cooling to below 25 ℃, wherein the cooling mode is natural cooling;
(3) Swirling the cooled centrifugal tube in the step (2) for 5s, then placing the centrifugal tube on a magnetic frame, standing for 4min until all magnetic beads are adsorbed to the wall of the centrifugal tube and the liquid in the centrifugal tube is clarified, absorbing the liquid, wherein the magnetic beads are not taken out when the liquid is absorbed, and in order to absorb all the liquid to the greatest extent, the residual liquid in the centrifugal tube is also absorbed, in the embodiment, the liquid is absorbed for the first time; instantly centrifuging the centrifuge tube, placing the centrifuge tube on a magnetic rack, and sucking and discarding residual liquid;
(4) And removing the magnetic frame, adding 50 mu L of DNA eluent into the centrifuge tube, wherein the DNA eluent is deionized water, blowing the magnetic beads in the centrifuge tube by using a pipette tip until the liquid in the centrifuge tube is uniformly distributed, and collecting the elution liquid in the centrifuge tube.
Example 7
The preparation of the lysis solution was the same as in example 1, except for the method of extracting nucleic acid from cervical swabs in the nucleic acid kit.
The kit for extracting the nucleic acid from the cervical swab comprises the cracking binding solution and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads and a preservative, the magnetic beads are used for adsorbing DNA, and the dosage of the magnetic beads is 400 mu g/ml.
The method for extracting nucleic acid from cervical swabs comprises the following steps:
(1) Adding 1000. Mu.L of the lysis binding solution and 25. Mu.L of the magnetic bead solution into a micro-centrifuge tube, mixing, adding 200. Mu.L of the cervical swab sample, mixing uniformly by vortex oscillation for 20s, and standing for 1min in a water bath at 95 ℃;
(2) Taking the centrifugal tube in the step (1) out of the water bath, standing, cooling to below 25 ℃, wherein the cooling mode is natural cooling or placing the centrifugal tube in cold water at the temperature of 25 ℃, and accelerating cooling;
(3) Swirling the centrifugal tube cooled in the step (2) for 15s, then placing the centrifugal tube on a magnetic frame, standing for 4min until all magnetic beads are adsorbed to the wall of the centrifugal tube and liquid in the centrifugal tube is clarified, absorbing and discarding liquid, wherein the magnetic beads are not taken out when the liquid is absorbed and discarded, and in order to completely absorb and discard the liquid as much as possible, residual liquid in the centrifugal tube is also required to be absorbed and discarded;
(4) And removing the magnetic frame, adding 50 mu L of DNA eluent into the centrifuge tube, wherein the DNA eluent is deionized water, blowing the magnetic beads in the centrifuge tube by using a pipette tip until the liquid in the centrifuge tube is uniformly distributed, and collecting the elution liquid in the centrifuge tube.
In the above examples, the DNA eluents were 50. Mu.L of deionized water for the same assay concentration, and in other embodiments, the volume of the DNA eluents may be 100. Mu.L or 40. Mu.L.
Comparative example 1
In this example, the difference from example 1 is that the lysis conjugate contains the following components in concentrations: sodium dodecyl sulfate 2%, octyl phenyl ether polyethylene glycol 1%, 4-hydroxyethyl piperazine ethanesulfonic acid 200mM, sodium chloride 10%, tris hydrochloride 100mM and EDTA 1mM (lacking polyoxyethylene lauryl ether sulfate sodium salt).
Comparative example 2
The commercially available kit was purchased and the extraction was checked according to the corresponding method.
The results of measuring the DNA concentration quality of the same batch of samples of examples 1 to 5 and comparative examples 1 to 2 are shown in Table 1
Table 1.Dna concentration quality test results.
| Concentration (ng/. Mu.L) | 260/280 | |
| Example 1 | 538.2 | 1.88 |
| Example 2 | 379.2 | 1.76 |
| Example 3 | 432.2 | 1.93 |
| Example 4 | 568.8 | 1.85 |
| Example 5 | 575.7 | 1.85 |
| Example 6 | 445.6 | 1.9 |
| Example 7 | 497.6 | 1.88 |
| Comparative example 1 | 267.0 | 1.9 |
| Comparative example 2 | 154.3 | 1.9 |
As can be seen from table 1, the optimal cleavage binding fluid formulation is: 2% sodium dodecyl sulfate, 1% polyoxyethylene octyl phenyl ether, 200mM 4-hydroxyethyl piperazine ethanesulfonic acid, 10% sodium chloride, 100mM tris (hydroxymethyl) aminomethane hydrochloride, 1mM EDTA and 2% sodium lauryl polyoxyethylene ether sulfate, the formula can greatly improve the mass concentration of nucleic acid extraction, and the use of sodium lauryl polyoxyethylene ether sulfate is indispensable.
In addition, in the method for extracting nucleic acid, the volume ratio of the magnetic bead solution to the lysis binding solution is 1.
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 "include", "including" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article, or terminal device including a series of elements includes not only those elements but also other elements not explicitly listed or inherent to such process, method, article, or terminal device. Without further limitation, an element defined by the phrases "comprising 8230; \8230;" or "comprising 8230; \8230;" does not exclude the presence of additional elements in a process, method, article, or terminal device that comprises the element. Further, herein, "greater than," "less than," "more than," and the like are understood to exclude the present numbers; the terms "above", "below", "within" and the like are to be understood as including the number.
Although the embodiments have been described, once the basic inventive concept is known, other variations and modifications can be made to the embodiments by those skilled in the art, so that the above embodiments are only examples of the present invention, and not intended to limit the scope of the present invention, and all equivalent structures or equivalent processes that can be used in the present specification or directly or indirectly applied to other related fields are encompassed by the present invention.
Claims (7)
1. A lysis binding solution for extracting nucleic acid from a cervical swab is characterized by comprising the following components in concentration:
2% of sodium dodecyl sulfate, 1% of polyethylene glycol octyl phenyl ether, 200mM 4-hydroxyethyl piperazine ethanesulfonic acid, 10% of sodium chloride, 100mM of tris (hydroxymethyl) aminomethane hydrochloride, 1mM of EDTA and 2% of sodium lauryl polyoxyethylene ether sulfate, wherein the pH value of the cleavage bonding solution is kept between 7.0 and 8.0.
2. A kit for extracting nucleic acid from cervical swab, comprising the lysis binding solution as defined in claim 1, and a magnetic bead solution, wherein the magnetic bead solution is formed by mixing magnetic beads for adsorbing DNA and a preservative, and the concentration of the magnetic bead solution is 200-400 μ g/ml.
3. The kit of claim 2, wherein the amount of magnetic beads is 300 μ g/ml.
4. A method for extracting nucleic acid from a cervical swab, comprising the steps of:
(1) Adding the lysis binding solution and the magnetic bead solution according to claim 1 into a centrifuge tube, uniformly mixing, then adding a cervical swab sample, uniformly mixing by vortex oscillation for 5-20s, and then standing for 1min in a water bath at 90 ℃, wherein the volume ratio of the magnetic bead solution to the lysis binding solution is 1;
(2) Taking out the centrifugal tube in the step (1) from the water bath, standing, and cooling to below 25 ℃;
(3) Whirling the centrifuge tube cooled in the step (2) for 5-15s, then placing the centrifuge tube on a magnetic frame until magnetic beads are completely adsorbed to the wall of the centrifuge tube and liquid in the centrifuge tube is clarified, and then absorbing the liquid;
(4) Removing the magnetic frame, adding DNA eluent into the centrifuge tube, wherein the volume ratio of the DNA eluent to the cervical swab sample is 1-2, blowing or suspending and oscillating the magnetic beads on the tube wall by using a pipette tip until the liquid in the centrifuge tube is uniformly distributed, and collecting the elution liquid in the centrifuge tube.
5. The method of claim 4, wherein in step (3), after the step of aspirating the liquid, the step of aspirating the residual liquid is performed in one of the following ways:
(1) Standing the centrifugal tube for 0.5-1min, allowing residual liquid to fall to the bottom of the centrifugal tube along the wall of the centrifugal tube, and sucking away the residual liquid;
(2) Instantly centrifuging the centrifuge tube, placing the centrifuge tube on a magnetic rack, and sucking and discarding residual liquid;
(3) The centrifuge tube was knocked so that the residual liquid on the wall of the centrifuge tube all fell to the bottom, and then the centrifuge tube was placed on a magnetic stand to suck away the residual liquid.
6. The method of claim 4, wherein in step (4), the volume ratio of the DNA eluate to the cervical swab sample is 1.
7. The method of claim 4, wherein the DNA eluent is deionized water, DEPC water or PCR mix.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110163794.3A CN112725413B (en) | 2021-02-05 | 2021-02-05 | Splitting and combining liquid, kit and method for extracting nucleic acid from cervical swab |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110163794.3A CN112725413B (en) | 2021-02-05 | 2021-02-05 | Splitting and combining liquid, kit and method for extracting nucleic acid from cervical swab |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112725413A CN112725413A (en) | 2021-04-30 |
| CN112725413B true CN112725413B (en) | 2022-10-14 |
Family
ID=75596104
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110163794.3A Active CN112725413B (en) | 2021-02-05 | 2021-02-05 | Splitting and combining liquid, kit and method for extracting nucleic acid from cervical swab |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112725413B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113652419A (en) * | 2021-08-17 | 2021-11-16 | 宁波上格石医用材料有限公司 | Cracking binding solution for nucleic acid extraction and extraction method |
| CN113755489A (en) * | 2021-10-13 | 2021-12-07 | 江苏溢纳生物科技有限公司 | Lysis binding solution for nucleic acid extraction and extraction method thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101481400B (en) * | 2009-01-07 | 2010-12-08 | 戴立忠 | Method for separating and purifying nucleic acid from sample of virus or cell containing nucleic acid and special magnetic separator therefor |
| DK2521780T3 (en) * | 2010-01-07 | 2017-12-04 | Bigtec Private Ltd | Process for Isolation of Nucleic Acids and Kits thereof |
| US9561166B2 (en) * | 2010-02-26 | 2017-02-07 | Hercules Incorporated | Polysaccharide products with improved performance and clarity in phosphate ester surfactant-based aqueous formulations and process for preparation |
| CN106675885A (en) * | 2016-12-30 | 2017-05-17 | 内蒙古河西航天科技发展有限公司 | High-alkalinity foam composite cleaning agent |
| AU2018286247B2 (en) * | 2017-06-14 | 2021-12-23 | Chia Tai Tianqing Pharmaceutical Group Co., Ltd. | Syk inhibitor and use method therefor |
| CN107759538B (en) * | 2017-10-30 | 2021-04-23 | 黑龙江八一农垦大学 | 2, 3-epoxy-2-nonane sulfone-5, 8-dimethoxy-1, 4-naphthoquinone, preparation method thereof and medicine containing same |
| CN112011594A (en) * | 2019-05-30 | 2020-12-01 | 苏州海狸生物医学工程有限公司 | Free nucleic acid extraction kit and extraction method thereof |
-
2021
- 2021-02-05 CN CN202110163794.3A patent/CN112725413B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN112725413A (en) | 2021-04-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112725413B (en) | Splitting and combining liquid, kit and method for extracting nucleic acid from cervical swab | |
| CN111057705B (en) | Kit for extracting free nucleic acid and use method | |
| CN108064262B (en) | Nucleic acid extraction device and method | |
| CN111484991B (en) | Kit for high-throughput full-automatic extraction of viral nucleic acid and extraction method | |
| CN104673783A (en) | Kit for extracting DNA/RNA of virus through magnetic bead method and using method | |
| CN107663521A (en) | Plasma free nucleic acid extraction kit and its application | |
| CN105349532A (en) | Method and kit for extracting free nucleic acid by using paramagnetic particle method | |
| JP2011505139A (en) | Method for isolating genomic DNA, RNA and protein from a single sample | |
| CN103930546A (en) | Rapid method for isolating extracellular nucleic acids | |
| CN102533724A (en) | Cell lysis reagent for extracting and purifying nucleic acids in biological samples | |
| CN106591297A (en) | Magnetic bead nucleic acid extraction method | |
| CN105695450A (en) | Magnetic-bead-process-based kit for extracting free DNAs (deoxyribonucleic acids) and application method thereof | |
| CN105524917A (en) | Kit for extracting blood genome DNA based on magnetic bead method and use method for kit | |
| CN107299097A (en) | A kind of micro-nucleic acid releasing agent, preparation method and applications | |
| CN109385418A (en) | A kind of method extracted for virus in animal specimen/bacterial nucleic acid and reagent | |
| CN107384856A (en) | A kind of method for preparing platelet lysates liquid | |
| CN106754890A (en) | The extracts kit and extracting method of a kind of viral RNA | |
| CN102676503A (en) | Method for quickly extracting DNA (deoxyribonucleic acid) | |
| CN110938624A (en) | Kit for extracting genome DNA and application thereof | |
| CN107119039A (en) | It is a kind of to organize not grinding the method for directly extracting nucleic acid | |
| CN117264945A (en) | Nucleic acid extraction kit based on magnetic bead method, application thereof and nucleic acid extraction method | |
| CN116590279A (en) | Kit for extracting plasma free DNA (deoxyribonucleic acid) based on hydroxyl magnetic beads and application method | |
| CN110951724A (en) | Method for rapidly extracting nucleic acid DNA and/or RNA by Trizol | |
| EP4253538A1 (en) | Method for isolating nucleic acid using binding buffer including compound having low or intermediate dielectric constant | |
| CN118109559A (en) | Kit for simply, conveniently and efficiently extracting plasma free DNA |
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 |