CN114426969A - Method for rapidly extracting nucleic acid for clinical sample detection - Google Patents
Method for rapidly extracting nucleic acid for clinical sample detection Download PDFInfo
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- YTRQFSDWAXHJCC-UHFFFAOYSA-N chloroform;phenol Chemical compound ClC(Cl)Cl.OC1=CC=CC=C1 YTRQFSDWAXHJCC-UHFFFAOYSA-N 0.000 description 1
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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- 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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Abstract
The invention relates to the technical field of clinical sample detection, in particular to a method for quickly extracting nucleic acid for clinical sample detection, which comprises the following steps: s1, taking 0.5-1 ml of clinical biological sample, adding the clinical biological sample into a test tube filled with mixed magnetic beads, standing for 12-15 min after uniform mixing, placing the test tube on a magnetic frame, standing for 3-5 min, then sending the test tube into a centrifuge for centrifugation, centrifuging for 3.5min under 12000rmp, and absorbing and discarding the supernatant; s2, adding 1.5ml of physiological saline into a test tube, shaking and scattering the sediment in a vortex oscillator, sending the sediment into a centrifuge for centrifugal treatment, centrifuging for 6-9 min under the condition of 12000rmp, sucking and discarding supernatant, adding 85-90 ul of first treatment fluid into the test tube by using a fluid-moving gun, and adding 15-20 ul of potassium acetate buffer solution after the solution in the test tube becomes clear.
Description
Technical Field
The invention relates to the technical field of clinical sample detection, in particular to a method for quickly extracting nucleic acid for clinical sample detection.
Background
Nucleic acid is a carrier of genetic information, is the material basis of gene expression, and no matter the research on nucleic acid structure or function is carried out, the required genome nucleic acid needs to be quickly and effectively separated and extracted from complex and various biological samples, and the quality and the integrity of the extracted nucleic acid directly influence the subsequent experimental results.
The major challenges for molecular diagnostics using nucleic acid amplification techniques are the pretreatment of nucleic acid-containing materials prior to amplification and the cumbersome nucleic acid extraction steps, and the silica-based and magnetic bead-based techniques that are currently widely used suffer from a number of drawbacks, including low efficiency (< 10%); there is also a need to optimize for specific target types, resulting in the extraction process of one sample type being generally incompatible with other sample types; the method using phenol chloroform has high recovery rate, but the application range is limited due to high toxicity of the components of the extracting solution; detergent-based methods are simple to solubilize, but require high temperature denaturation (95 ℃) and are not suitable for single-step RT-PCR because reverse transcriptase is not thermostable.
The existing method for extracting nucleic acid needs repeated centrifugation, has complex extraction steps, consumes long time, needs specific equipment and cannot extract nucleic acid quickly.
In summary, the present invention provides a rapid nucleic acid extraction method for clinical specimen detection to solve this problem.
Disclosure of Invention
The present invention aims to provide a method for rapidly extracting nucleic acid for clinical specimen detection, so as to solve the problems in the background art.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for rapidly extracting nucleic acid for clinical sample detection comprises the following steps:
s1, taking 0.5-1 ml of clinical biological sample, adding the clinical biological sample into a test tube filled with mixed magnetic beads, standing for 12-15 min after uniform mixing, placing the test tube on a magnetic frame, standing for 3-5 min, then sending the test tube into a centrifuge for centrifugation, centrifuging for 3.5min under 12000rmp, and absorbing and discarding the supernatant;
s2, adding 1.5ml of physiological saline into a test tube, shaking and scattering the sediment in a vortex oscillator, sending the test tube into a centrifuge for centrifugal treatment, centrifuging for 6min to 9min under the condition of 12000rmp, sucking and discarding supernatant, adding 85ul to 90ul of first treatment fluid into the test tube by using a fluid-moving gun, adding 15ul to 20ul of potassium acetate buffer solution after the solution in the test tube becomes clear, mixing uniformly, sending the test tube into the centrifuge again for centrifugal treatment after white flocculent sediment appears in the test tube, centrifuging for 6.5min to 8.5min under the condition of 12000rmp, sucking and discarding supernatant, adding washing fluid into the test tube, mixing uniformly, placing the test tube on a magnetic frame for 5.5min to 6min, and collecting to obtain a clinical biological sample nucleic acid solution;
s3, sending the nucleic acid solution of the clinical biological sample into a collecting tube filled with the extracting solution, mixing uniformly, dipping the adsorption film into the extracting solution, standing for 15min-20min, taking out the adsorption film, slowly washing the adsorption film by using eluent, then placing the adsorption film into the PCR reaction solution, adjusting the pH of the PCR reaction solution to 8.4, and releasing the nucleic acid from the adsorption film to the PCR reaction solution.
In a preferred embodiment of the present invention, the mixed magnetic beads in S1 are prepared by mixing immunomagnetic beads and nucleic acid-extracting magnetic beads, wherein the immunomagnetic beads are MNP-abs obtained by bonding a virus antibody or receptor to a hydroxyl group, a carboxyl group, an amino group or an epoxy group on the surface of a magnetic microsphere, the nucleic acid-extracting magnetic beads are silica-hydroxyl magnetic beads, carboxyl magnetic beads or other magnetic beads capable of binding virus nucleic acid under specific conditions, the virus antibody includes an outer membrane protein of enveloped virus, a coat protein antibody without enveloped virus or a nucleocapsid protein antibody, the outer membrane protein antibody of enveloped virus is an antibody against F protein, M protein, N protein or S protein, and the virus receptor is a monomer or a multi-molecule complex specifically bound to virus.
In a preferred embodiment of the present invention, when the first treatment solution is added using a pipette in S2, the pipette is used to gently blow the precipitated cells in the test tube, and the test tube is slowly shaken by closing the tube cover without violent shaking.
In a preferred embodiment of the present invention, the first treatment solution in S2 is prepared by mixing Chelex-100, MNaOH, SDS and physiological saline, wherein the concentration of Chelex-100 is 11.5% to 12%, the concentration of NaOH is 0.45M to 0.48M, the concentration of SDS is 5.5% to 5.8%, and the pH of the first treatment solution is 13.5 to 13.9.
In a preferred embodiment of the present invention, the pH of the potassium acetate buffer solution in S2 is 5.8-5.9.
As a preferable scheme of the present invention, the specific operation steps of uniformly mixing in S2 are: the tube mouth was closed, the tube was inverted several times and mixed well and the tube bottom was flicked several times with the finger.
As a preferable scheme of the invention, the extracting solution in the S3 is prepared by mixing guanidinium isothiocyanate, TritonX100, Tween-20, proteinase K, MNaCl, MEDTA, Tween-20 and SDS, wherein the concentration of the guanidinium isothiocyanate is 2.2M-2.3M, the concentration of the TritonX100 is 0.52 wt%, the concentration of the Tween-20 is 1.3 wt%, the concentration of the proteinase K is 61 mu g/mL, the concentration of the MNaCl is 151M, the concentration of the MEDTA is 12M, the concentration of the SDS is 0.55%, and the pH value of the extracting solution is 8.1.
In a preferred embodiment of the present invention, the adsorption film in S3 is a cellulose film treated with an anionic resin containing a quaternary ammonium salt.
As a preferable scheme of the invention, the eluent in S3 is 0.12 wt% Tween-20 Tris buffer, and the pH value of the eluent is 8.2.
As a preferable scheme of the invention, the washing solution in S2 is prepared by mixing 5.5M guanidine hydrochloride, 56% absolute ethyl alcohol and 80 mug/mL proteinase K, and the pH value is 6-7.
Compared with the prior art, the invention has the beneficial effects that:
1. in the invention, a clinical biological sample is added into a test tube filled with mixed magnetic beads, the test tube is evenly mixed and then sent into a centrifuge for centrifugal treatment, supernatant is absorbed and discarded, normal saline is added into the test tube, the test tube is shaken in a vortex oscillator to disperse and precipitate, the test tube is sent into the centrifuge for centrifugal treatment, the supernatant is absorbed and discarded, a first treatment solution and a potassium acetate buffer solution are added into the test tube by using a liquid-moving gun, the test tube is evenly mixed, after white flocculent precipitate appears in the test tube, the test tube is sent into the centrifuge again for centrifugal treatment, the supernatant is taken and added into the test tube, washing liquid is collected to obtain a clinical biological sample nucleic acid solution, the clinical biological sample nucleic acid solution is sent into a collecting tube filled with extracting solution, after the uniform mixing, an adsorption film is soaked into the extracting solution, the adsorption film is taken out, the adsorption film is slowly cleaned by the washing liquid, and then the adsorption film is placed into PCR reaction liquid, the adsorption membrane releases nucleic acid to PCR reaction liquid, the immunomagnetic beads and the nucleic acid extraction magnetic beads can carry out specific adsorption on the nucleic acid and are eluted in washing liquid, thereby achieving the purpose of extracting and purifying the nucleic acid, the specific adsorption rate is high, the recovery rate of virus is high, the repeatability is strong, the automatic operation steps are few, the cost is low, meanwhile, the alkaline first treatment liquid is adopted to crack a sample, the cell membrane is broken, protein in cells is combined with Sodium Dodecyl Sulfate (SDS), sodium ions and potassium ions in the SDS are replaced to generate water-insoluble Potassium Dodecyl Sulfonate (PDS), thereby achieving the purpose of separating the nucleic acid, the whole extraction process takes shorter time, the extraction time is greatly shortened, the nucleic acid with negative electricity is adsorbed to a cellulose membrane treated by anion resin containing quaternary ammonium salt under the condition of weak alkalinity, and then the elution is carried out by elution liquid, the method has the advantages of rapid extraction process, less impurities, less limitation on temperature, and difficult dissociation of nucleic acid, thereby rapidly extracting nucleic acid samples.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments, and based on the embodiments of the present invention, all other embodiments obtained by a person skilled in the art without making creative efforts belong to the protection scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, and the terms used herein in the specification of the present invention are for the purpose of describing particular embodiments only and are not intended to limit the present invention, and the term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The invention provides a technical scheme that:
a method for rapidly extracting nucleic acid for clinical sample detection comprises the following steps:
s1, taking 0.5ml to 1ml of clinical biological sample, adding the clinical biological sample into a test tube filled with mixed magnetic beads, uniformly mixing, standing for 12min to 15min, placing the test tube on a magnetic frame, standing for 3min to 5min, then sending the test tube into a centrifugal machine for centrifugal treatment, centrifuging for 3.5min under the condition of 12000rmp, and sucking and removing supernatant;
s2, adding 1.5ml of physiological saline into a test tube, shaking and scattering the sediment in a vortex oscillator, sending the test tube into a centrifuge for centrifugal treatment, centrifuging for 6min to 9min under the condition of 12000rmp, sucking and discarding supernatant, adding 85ul to 90ul of first treatment fluid into the test tube by using a fluid-moving gun, adding 15ul to 20ul of potassium acetate buffer solution after the solution in the test tube becomes clear, mixing uniformly, sending the test tube into the centrifuge again for centrifugal treatment after white flocculent sediment appears in the test tube, centrifuging for 6.5min to 8.5min under the condition of 12000rmp, sucking and discarding supernatant, adding washing fluid into the test tube, mixing uniformly, placing the test tube on a magnetic frame for 5.5min to 6min, and collecting to obtain a clinical biological sample nucleic acid solution;
s3, sending the nucleic acid solution of the clinical biological sample into a collecting tube filled with the extracting solution, mixing uniformly, dipping the adsorption film into the extracting solution, standing for 15min-20min, taking out the adsorption film, slowly washing the adsorption film by using eluent, then placing the adsorption film into the PCR reaction solution, adjusting the pH of the PCR reaction solution to 8.4, and releasing the nucleic acid from the adsorption film to the PCR reaction solution.
Further, the mixed magnetic beads in S1 are prepared by mixing immunomagnetic beads and nucleic acid-extracting magnetic beads, the immunomagnetic beads are MNP-abs obtained by bonding virus antibodies or receptors with hydroxyl, carboxyl, amino or epoxy groups on the surface of the magnetic microspheres, the nucleic acid-extracting magnetic beads are silicon hydroxyl magnetic beads, carboxyl magnetic beads or other magnetic beads capable of binding virus nucleic acids under specific conditions, the virus antibodies include outer membrane proteins of enveloped viruses, coat protein antibodies or nucleocapsid protein antibodies without enveloped viruses, the outer membrane protein antibodies of enveloped viruses are antibodies of F protein, M protein, N protein or S protein, and the virus receptors are monomer or multi-molecule composites specifically bound to viruses.
Further, when the first treatment solution is added by using the pipette in S2, the pipette is required to gently blow off the precipitated cells in the test tube, and the test tube is slowly shaken by closing the tube cover without violent shaking.
Furthermore, the first treatment fluid in the S2 is prepared by mixing Chelex-100, MNaOH, SDS and normal saline, the concentration of the Chelex-100 is 11.5-12%, the concentration of NaOH is 0.45-0.48M, the concentration of the SDS is 5.5-5.8%, and the pH of the first treatment fluid is 13.5-13.9.
Further, the pH of the potassium acetate buffer solution in the S2 is 5.8-5.9.
Further, the specific operation steps of uniformly mixing in S2 are as follows: the tube mouth was closed, the tube was inverted several times and mixed well and the tube bottom was flicked several times with the finger.
Furthermore, the extracting solution in the S3 is prepared by mixing guanidinium isothiocyanate, TritonX100, Tween-20, proteinase K, MNaCl, MEDTA, Tween-20 and SDS, wherein the concentration of the guanidinium isothiocyanate is 2.2M-2.3M, the concentration of the TritonX100 is 0.52 wt%, the concentration of the Tween-20 is 1.3 wt%, the concentration of the proteinase K is 61 mu g/mL, the concentration of the MNaCl is 151M, the concentration of the MEDTA is 12M, the concentration of the SDS is 0.55%, and the pH value of the extracting solution is 8.1.
Further, the adsorption film in S3 is a cellulose film treated with an anionic resin containing a quaternary ammonium salt.
Further, the eluent in S3 is 0.12 wt% Tween-20 Tris buffer, and the pH value of the eluent is 8.2.
Further, the washing solution in S2 is prepared by mixing 5.5M guanidine hydrochloride, 56% absolute ethyl alcohol and 80 μ g/mL proteinase K, and the pH value is 6-7.
The specific implementation case is as follows:
taking 1ml of a clinical biological sample, adding the clinical biological sample into a test tube filled with mixed magnetic beads, wherein the mixed magnetic beads are prepared by mixing immunomagnetic beads and nucleic acid extraction magnetic beads, the immunomagnetic beads are MNP-Ab obtained by bonding virus antibodies or receptors and the like with hydroxyl, carboxyl, amino or epoxy groups on the surfaces of magnetic microspheres, the nucleic acid extraction magnetic beads are silicon hydroxyl magnetic beads, carboxyl magnetic beads or other magnetic beads capable of combining virus nucleic acid under specific conditions, standing for 12min after uniform mixing, placing the test tube on a magnetic frame, standing for 3min, sending the test tube into a centrifugal machine for centrifugal treatment, centrifuging for 3.5min under the condition of 12000rmp, and sucking and discarding supernatant;
adding 1.5ml of normal saline into a test tube, shaking and scattering the sediment in a vortex oscillator, sending the test tube into a centrifuge for centrifugal treatment, centrifuging for 6min under the condition of 12000rmp, sucking and removing supernatant, adding 85ul of first treatment fluid into the test tube by using a fluid transfer gun, mixing the first treatment fluid with Chelex-100, MNaOH, SDS and normal saline, wherein the pH of the first treatment fluid is 13.5-, gently blowing off the precipitated cells in the test tube by using the fluid transfer gun, tightly covering a tube cover to slowly shake the test tube without violent shaking, adding 15ul of potassium acetate buffer solution after the solution in the test tube becomes clear, covering a tube opening tightly, turning the test tube over for a plurality of times, mixing the test tube evenly, flicking the bottom of the test tube for a plurality of times by using a finger, mixing the test tube evenly, sending the test tube into the centrifuge again for centrifugal treatment after white sediment appears in the test tube, centrifuging for 8.5min under the condition of 12000rmp, absorbing and removing the supernatant, adding a washing solution into the test tube, wherein the washing solution is prepared by mixing 5.5M guanidine hydrochloride, 56% absolute ethyl alcohol and 80 mug/mL proteinase K, the pH value is 6.5, after the mixture is uniform, placing the mixture on a magnetic frame for 6min, and collecting eluent to obtain a clinical biological sample nucleic acid solution;
feeding nucleic acid solution of clinical biological sample into a collecting tube containing extracting solution, wherein the extracting solution in S3 is prepared by mixing guanidinium isothiocyanate with concentration of 2.2M, TritonX100 with concentration of 0.52 wt%, Tween-20 with concentration of 1.3 wt%, proteinase K with concentration of 61 mug/mL, MNaCl with concentration of 151M, MEDTA with concentration of 12M and SDS with concentration of 0.55%, the pH value of the extracting solution is 8.1, mixing uniformly, soaking an adsorption membrane in the extraction solution, wherein the adsorption membrane is a cellulose membrane treated by quaternary ammonium salt-containing anion resin, standing for 15-20 min, taking out the adsorption membrane, slowly washing the adsorption membrane by using eluent, the eluent in S3 is 0.12 wt% of Tween-20 Tris buffer solution, the pH value of the eluent is 8.2, placing the adsorption membrane in PCR reaction solution, adjusting the pH value of the PCR reaction solution to 8.4, and releasing nucleic acid from the adsorption membrane to the PCR reaction solution.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A method for rapidly extracting nucleic acid for clinical sample detection is characterized by comprising the following steps:
s1, taking 0.5ml to 1ml of clinical biological sample, adding the clinical biological sample into a test tube filled with mixed magnetic beads, uniformly mixing, standing for 12min to 15min, placing the test tube on a magnetic frame, standing for 3min to 5min, then sending the test tube into a centrifugal machine for centrifugal treatment, centrifuging for 3.5min under the condition of 12000rmp, and sucking and removing supernatant;
s2, adding 1.5ml of physiological saline into a test tube, shaking and scattering the sediment in a vortex oscillator, sending the test tube into a centrifuge for centrifugal treatment, centrifuging for 6min to 9min under the condition of 12000rmp, sucking and discarding supernatant, adding 85ul to 90ul of first treatment fluid into the test tube by using a fluid-moving gun, adding 15ul to 20ul of potassium acetate buffer solution after the solution in the test tube becomes clear, mixing uniformly, sending the test tube into the centrifuge again for centrifugal treatment after white flocculent sediment appears in the test tube, centrifuging for 6.5min to 8.5min under the condition of 12000rmp, sucking and discarding supernatant, adding washing fluid into the test tube, mixing uniformly, placing the test tube on a magnetic frame for 5.5min to 6min, and collecting to obtain a clinical biological sample nucleic acid solution;
s3, sending the nucleic acid solution of the clinical biological sample into a collecting tube filled with the extracting solution, mixing uniformly, dipping the adsorption film into the extracting solution, standing for 15min-20min, taking out the adsorption film, slowly washing the adsorption film by using eluent, then placing the adsorption film into the PCR reaction solution, adjusting the pH of the PCR reaction solution to 8.4, and releasing the nucleic acid from the adsorption film to the PCR reaction solution.
2. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the mixed magnetic beads in the S1 are prepared by mixing immunomagnetic beads and nucleic acid extraction magnetic beads, wherein the immunomagnetic beads are MNP-Abs obtained by bonding virus antibodies or receptors and the like with hydroxyl, carboxyl, amino or epoxy groups on the surfaces of the magnetic microspheres, the nucleic acid extraction magnetic beads are silicon hydroxyl magnetic beads, carboxyl magnetic beads or other magnetic beads capable of combining virus nucleic acids under specific conditions, the virus antibodies comprise envelope proteins of enveloped viruses, coat protein antibodies without the enveloped viruses or nucleocapsid protein antibodies, the envelope protein antibodies of the enveloped viruses are antibodies of F proteins, M proteins, N proteins or S proteins, and the virus receptors are monomers or multi-molecule compounds specifically combined with the viruses.
3. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: when the first treatment solution is added by using the pipette in S2, the pipette is used to gently blow the precipitated cells in the test tube, and the tube cover is closed to slowly shake the test tube without violent shaking.
4. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the first treatment fluid in the S2 is prepared by mixing Chelex-100, MNaOH, SDS and normal saline, wherein the concentration of the Chelex-100 is 11.5-12%, the concentration of NaOH is 0.45-0.48M, the concentration of SDS is 5.5-5.8%, and the pH value of the first treatment fluid is 13.5-13.9.
5. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the pH value of the potassium acetate buffer solution in the S2 is 5.8-5.9.
6. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the specific operation steps of uniformly mixing in the step S2 are as follows: the tube mouth was closed, the tube was inverted several times and mixed well and the tube bottom was flicked several times with the finger.
7. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the extracting solution in the S3 is prepared by mixing guanidinium isothiocyanate, TritonX100, Tween-20, proteinase K, MNaCl, MEDTA, Tween-20 and SDS, wherein the concentration of the guanidinium isothiocyanate is 2.2M-2.3M, the concentration of the TritonX100 is 0.52 wt%, the concentration of the Tween-20 is 1.3 wt%, the concentration of the proteinase K is 61 mu g/mL, the concentration of the MNaCl is 151M, the concentration of the MEDTA is 12M, the concentration of the SDS is 0.55%, and the pH value of the extracting solution is 8.1.
8. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the adsorption film in S3 is a cellulose film treated by anion resin containing quaternary ammonium salt.
9. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the eluent in S3 is 0.12 wt% Tween-20 Tris buffer, and the pH value of the eluent is 8.2.
10. The method for rapidly extracting nucleic acid for clinical specimen detection according to claim 1, wherein: the washing solution in the S2 is prepared by mixing 5.5M guanidine hydrochloride, 56% absolute ethyl alcohol and 80 mu g/mL proteinase K, and the pH value is 6-7.
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| CN117861627A (en) * | 2024-01-03 | 2024-04-12 | 湖北擎科生物科技有限公司 | Potassium ion microsphere, plasmid extraction reagent and application |
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