CN102115739B - Method, reagent and kit for separating nucleic acids - Google Patents
Method, reagent and kit for separating nucleic acids Download PDFInfo
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- CN102115739B CN102115739B CN200910217168.7A CN200910217168A CN102115739B CN 102115739 B CN102115739 B CN 102115739B CN 200910217168 A CN200910217168 A CN 200910217168A CN 102115739 B CN102115739 B CN 102115739B
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Abstract
The invention provides a simple and quick method for separating nucleic acids, which comprises the following steps: providing a nucleic acid separation reagent, mixing the nucleic acid separation reagent with a biological sample containing one or multiple nucleic acids, and separating the nucleic acids from the biological sample by one step under a proper reaction condition without pretreating the sample. The reagent contains 1-40 wt% of polyethylene glycol (PEG), and/or higher than 30 wt% of low-molecular-weight alcohol, salt and surfactant. The nucleic acids can be combined with a solid-phase vector by changing the solubility of the nucleic acids, thereby separating the nucleic acids. In addition, the invention also provides a reagent and kit for separating nucleic acids.
Description
Technical field
The invention relates to a kind of method of isolating nucleic acid, and be particularly to a kind of separate nucleic acid method and separate nucleic acid reagent of one step.
Background technology
In recent years, molecular diagnosis, medicine Genomics, translate the rise such as medical science and criminal identification, the demand of various foranalysis of nucleic acids also increases rapidly, but, no matter carry out which kind of foranalysis of nucleic acids, first necessarily require highly purified nucleic acid, and separate nucleic acid method easily and fast.
In the time analyzing, because the time that test substance exists in liquid sample is very of short duration, and the impurity in sample can affect the accuracy detecting, therefore develop a kind of isolation technique of nucleic acid, utilize a solid phase carrier bind nucleic acid, or using pipe string internal wall as solid phase carrier, then add microballon to tubing string, again microballon is separated after utilizing microballon bind nucleic acid.In addition, also developing the method for utilizing magnetic bead isolating nucleic acid at present, is mainly to utilize magnetic bead bind nucleic acid, and recycling magnetic is separated in connection with the magnetic bead of nucleic acid.
Along with the demand of separate nucleic acid is more and more frequent, the sample of action required source is also more and more extensive, because magnetic separation technique does not need centrifugal or the step such as vacuumizes, therefore can save treatment time and cost.In addition, the purge process of magnetic resolution is quick, easy to operate, therefore magnetic resolution develops into main separate nucleic acid method gradually at present.
At present common separate nucleic acid technology is mainly first to process complex samples nucleic acid is disengaged on the market, recycling solid phase is in conjunction with isolating nucleic acid, the Chaotropic Salt (chaotropic salt) of solid phase separate part application Boom Patent (No. 5234809 patent of the U.S.) is adsorbed to the nucleic acid in sample on silicon materials (Silica), through after cleaning step, re-use punching and carry (elution, wash-out) damping fluid rinses the nucleic acid on silicon materials get off, and reaches the effect of isolating nucleic acid.In addition, No. 6274386 patent of the U.S. is the magnetic solid phase carrier absorption nucleic acid of the siliceous surfacing of application.
But, anyway, 5 steps such as all separate nucleic acid methods all need through dissolving (1ysis), extraction, precipitation, clean, punching is carried, its time spending is long and easily pollute.Therefore, the invention provides one fast and separate nucleic acid method and reagent (composition) easily, can save tradition multi-step separate nucleic acid method complicated and consuming time.
Summary of the invention
The present invention is to provide a kind of method of isolating nucleic acid, after the method comprises that the biological specimen that a separation agent and is contained to nucleic acid mixes, described separation agent can make nucleic acid in biological specimen, separate and be bonded on a solid phase carrier, do not need the Sample pretreatment can single stage isolating nucleic acid, it is characterized in that: the separation agent of use comprises the polyoxyethylene glycol of 1～40wt% (PEG) and/or is greater than lower molecular weight alcohols, salt and the tensio-active agent of 30wt%, is bonded to solid phase carrier by changing nucleic acid solubleness.
The present invention separately provides a kind of reagent of isolating nucleic acid, and this reagent comprises the polyoxyethylene glycol (PEG) of 1～40wt% and/or is greater than lower molecular weight alcohols, a salt, a tensio-active agent and a solid phase carrier of 30wt%.
The present invention also provides a kind of cover group of isolating nucleic acid, this cover group comprises a separation agent, its polyoxyethylene glycol that contains 1～40wt% (PEG) and/or be greater than lower molecular weight alcohols, salt, tensio-active agent, solid phase carrier and/or the proteolytic enzyme of 30wt%, and a container.
Preferably, in the present invention:
The molecular weight of described lower molecular weight alcohols is 30～100g/mol.
Described lower molecular weight alcohols is ethanol.
The weight concentration of described lower molecular weight alcohols is 40～60%.
The weight concentration of described polyoxyethylene glycol is 5～20%.
Described solid phase carrier comprises metal, metal oxide, silicide or polymkeric substance.
Described solid phase carrier comprises magnetic substance.
Described salt comprises alkali metal group, alkaline earths and/or ammonium salt halogenide.
Described tensio-active agent comprises Tween-20, Triton X-100, NP40, Brij35 or CHAPS.
Described reagent also comprises a proteolytic enzyme.
Described reagent also comprises a carbohydrate.Wherein said reagent comprises a polyose or polysaccharide derivatives.Wherein said polyose comprises glycogen, starch, oligose, dextran, Mierocrystalline cellulose, agarose, chitosan, mucopolysaccharide, peptidoglycan.
The hybrid reaction temperature of described reagent and biological specimen is approximately 50 DEG C～80 DEG C.
The mixing acid base number of described biological specimen and reagent is pH 6～9.
Described biological specimen includes one or more nucleic acid.
Described biological specimen includes cellular material.
Described biological specimen comprises prokaryotic organism, fungi, virus, cell, blood, amniotic fluid, cerebrospinal fluid, or from tissue and the tissue juice thereof of skin, muscle, conjunctiva, placenta or intestines and stomach.
Method of the present invention also can be included in after described biological specimen and reagent mix, carries out a wash procedure.The method is also included in after described wash procedure, carries out a punching and puies forward program.
Cover group of the present invention also can comprise that a cleaning buffer solution and/or a punching carry damping fluid.Wherein said cleaning buffer solution can be removed salt and protein.Damping fluid is carried in wherein said punching can be by nucleic acid by rinsing on solid phase carrier.
In cover group of the present invention, described container is for splendid attire separation agent.
For above and other objects of the present invention, feature and advantage can be become apparent, below spy enumerates preferred embodiment, and coordinates accompanying drawing, elaborates.
Brief description of the drawings
Fig. 1 is the electrophoretic analysis of DNA agar-agar.
Embodiment
The present invention is to provide a kind of method of separation and/or purification of nucleic acid.In the method for the invention, a separation agent is mixed with a biological specimen and react after separable go out nucleic acid in biological specimen.
In one embodiment, separation agent of the present invention comprises the polyoxyethylene glycol (PEG) of 1～40wt%, is preferably the polyoxyethylene glycol of 5～20wt%.
In another embodiment, separation agent of the present invention comprises the lower molecular weight alcohols that is greater than 30wt%, is preferably the lower molecular weight alcohols of 40～60wt%, and wherein the molecular weight of lower molecular weight alcohols is about 30～100g/mol.In one embodiment, lower molecular weight alcohols is ethanol.
In another embodiment, separation agent of the present invention can comprise polyoxyethylene glycol and lower molecular weight alcohols simultaneously.
Separation agent of the present invention more comprises a salt, a tensio-active agent, a proteolytic enzyme and a solid phase carrier.
Salt of the present invention can be any salt that is applicable to molecular biosciences experiment, comprises an alkali metal group or alkaline earths and/or amine salt, for example, and Li
+, Na
+, K
+, Rb
+, Cs
+, Fr
+, Gu
+(guanidine), Be
2+, Mg
2+, Ca
2+, Sr
2+, Ba
2+, Ra
2+and/or NH
4 +.In one embodiment, salt of the present invention can comprise a haloid element, is in particular Cl
-.In general, conventional salt is Gu-HCl (guanidinehydrochloride), LiCl, NaCl, Tris-HCl, Gu-SCN (guanidine thiocyanate) or its analogue.
Tensio-active agent of the present invention can be ionogenic surfactant, amphoterics or nonionic surface active agent, wherein be preferably nonionic surface active agent and amphoterics, there is no particular restriction for the kind of nonionic surface active agent, be preferably the nonionic surface active agent of polyoxyethylene class (polyoxyethylene), as Tween-20 (polysorbas20), Triton X-100 (Triton X-100), NP40 (polyoxyethylene nonylphenol ether) or Brij35 (polyoxyethylene laurel ether) etc.Usage quantity or the concentration of nonionogenic tenside do not limit, for example, can be 0.01% to 40%, be preferably 5% to 20%, there is no particular restriction for the kind of amphoterics, be preferably CHAPS (3-{ (3-cholamidopropyl) dimethylammonio}-propanesulfonate, 3-[3-(courage amido propyl) dimethylamino] propanesulfonic acid inner salt), usage quantity or the concentration of amphoterics do not limit, for example can be 0.01% to 20%, be preferably 1% to 10%.
Proteolytic enzyme of the present invention, can be any material with protein hydrolysate effect, such as thermolysin, stomach en-, trypsinase, bromeline (bromelain), alkaline hydrolysis proteolytic enzyme (Alcalase), Proteinase K (Protease K), flavor protease (Flavorzyme) or the beneficial auspicious proteolytic enzyme of alkalescence (Esperase) etc., be preferably Proteinase K (Protease K).
Separation agent of the present invention can add polyose (polysaccharides), comprise glycogen (glycogen), starch (starch), oligose (oligosaccharides), dextran (dextran), Mierocrystalline cellulose (cellulose), agarose (agarose), chitosan (chitosan), mucopolysaccharide (mucopolysaccharides) or peptidoglycan (peptidoglycan), the binding ability of small molecules nucleic acid and solid phase carrier be can increase, nucleic acid productive rate and purity improved.
Biological specimen of the present invention is a kind of biological specimen that contains nucleic acid.There is no particular restriction for biological specimen of the present invention, can be any sample with nucleic acid, karyomit(e) and/or plastid.In one embodiment, biological specimen of the present invention can be fungi, prokaryotic organism (bacterium), virus, zooblast or vegetable cell.In another embodiment, biological specimen of the present invention can be blood, amniotic fluid, cerebrospinal fluid or the tissue juice from skin, muscle, oral mucosa, placenta, intestines and stomach or other organ of animal.It should be noted, biological specimen of the present invention need, via any pre-treatment, can directly not react with separation agent, can isolate needed nucleic acid.
Nucleic acid of the present invention can be DNA or the RNA molecule of any natural or synthetic, comprises single strand dna, double chain DNA molecule, RNA, LNA, PNA, nucleic acid-protein complex and/or nucleic acid-carbohydrate complex body etc.
After separation agent of the present invention mixes with biological specimen, under a suitable condition, carry out a reactions steps.The temperature of reactions steps can be room temperature, 50 DEG C～80 DEG C, is preferably 60～70 DEG C; PH value can be 6～9, is preferably 7～8, and the reaction times can be 5～30 minutes, is preferably 10～20 minutes, and the best is 15 minutes.It should be noted, biomaterial can directly react with separation agent of the present invention, and must not carry out any pre-treatment.
Add after separation agent, if biological specimen is complex biological sample or a whole blood corpse or other object for laboratory examination and chemical testing that contains cell kenel, separation agent can make cell rupture disengage nucleic acid, and nucleic acid in sample can be separated because solubleness changes, do not need to carry out in advance that sample is centrifugal, the molten pre-treatment step such as split of cellular segregation, cell.Those skilled in the art can select any applicable method to be further purified nucleic acid, for example: the nucleic acid of separation can utilize alcohol to clean to obtain more highly purified nucleic acid.
In another embodiment, after the nucleic acid of separation can be combined with solid phase carrier, then punching puts forward.Separation agent of the present invention, except meeting changes the solubleness of nucleic acid, also can change the structure of nucleic acid, makes nucleic acid be easier to paste on solid phase carrier, to reach the object of separation.
Solid phase carrier of the present invention can be a kind of magnetic substance, metal, metal oxide, silicide, polymkeric substance.The surface of solid phase carrier can be modified by chemical bond.In a preferred embodiment, the surface of solid phase carrier has carbohydrate (hydrocarbon polymer), for example, and Mierocrystalline cellulose, soluble cotton, dextran or its analogue.In another embodiment, the surface of solid phase carrier has a hydrophobic grouping, for example, and C
1～C
30alkyl or C
5～C
30aryl etc.In another embodiment, the surface of solid phase carrier can have a hydrophilic radical, for example, and hydroxyl, carbonyl, carboxyl, ester class, amido, sulfenyl etc.
In addition,, after reactions steps, can sequentially carry out a wash procedure and punching and put forward program.
Wash procedure comprises and adds a cleaning buffer solution, to remove unnecessary salt and protein.Cleaning buffer solution of the present invention can be any applicable cleaning buffer solution, and it is mainly the protein, glycan class, lipid or the cell debris that remove on nucleic acid.In general, cleaning buffer solution can comprise alcohols and salt, and alcohols can be ethanol or polyoxyethylene glycol.Salt can be lower than the sodium-chlor of 3M or its analogue.In general, the volume of cleaning buffer solution and wash number, to be enough to removal of impurity (protein, glycan class, lipid or cell debris), are good but do not affect nucleic acid output.
After wash procedure, can carry out a punching and put forward program.Punching is put forward program and is comprised that adding a punching to carry damping fluid is put forward nucleic acid by solid phase carrier upper punch.Punching is carried damping fluid and be can be water, TE, TAE or TBE or its analogue.
The method of separation of the present invention and/or purification of nucleic acid is used single agents, completes with one step, can obtain fast, easily the nucleic acid of high purity and high yield.The more important thing is, biological specimen need, via any pre-treatment, can directly not react with separation agent, can isolate needed nucleic acid.
The present invention separately provides a kind of separation agent of nucleic acid, at least comprises the polyoxyethylene glycol (PEG) of 1～40wt% and/or is greater than the lower molecular weight alcohols of 30wt%.
Separation agent of the present invention can further comprise above-mentioned salt, tensio-active agent, solid phase carrier and/or proteolytic enzyme.
The present invention also provides a kind of cover group of isolating nucleic acid, comprise a separation agent, its polyoxyethylene glycol that contains 1～40wt% (PEG) and/or be greater than lower molecular weight alcohols, salt, tensio-active agent and solid phase carrier, a cleaning buffer solution, a container of 30wt%, and working instructions.
[embodiment]
1. the separate nucleic acid of whole blood
PEG (0%, 5%, 10%, 15%, 20%, 30%), salt that the separation agent using in this embodiment contains different concns are that 3M guanidine hydrochloride (Gu-HCl), tensio-active agent are the magnetic solid phase carrier of 5%Tween-20 and 0.5 μ g, and contain proteolytic enzyme and 6M urea.
The separation agent of the present invention of getting 720 μ l directly mixes with the human whole blood of 200 μ l, at 56 DEG C, react after 10 minutes, separate nucleic acid can be bonded on magnetic solid phase carrier, remove after the aqueous solution, washed with cleaning buffer solution, finally add 200 μ l aseptic deionized waters to rush the nucleic acid that dike after 10 minutes puts forward magnetic solid phase carrier upper punch and analyze.The cleaning buffer solution that this embodiment uses is 70%EtOH.
As shown in Table 1, separation method of the present invention does not need the red blood corpuscle in pre-treatment whole blood sample, can be effectively directly by DNA by separating in whole blood, can in the human whole blood of 200 μ l, isolate approximately 1～3 μ g DNA, purity (A
260/ A
280) be about 1.3～1.6.
The effect that table one, DNA separate
| PEG concentration (%) | (μ g) for DNA fractional dose | DNA purity (A 260/A 280) |
| 0 | 0.73 | 1.31 |
| 5 | 1.68 | 1.55 |
| 10 | 3.21 | 1.62 |
| 15 | 2.85 | 1.46 |
| 20 | 2.12 | 1.35 |
| 30 | 1.03 | 1.26 |
In addition, ethanol (40%, 50% and 60%), the salt that uses different concns in separation agent is that 3M guanidine hydrochloride (Gu-HCl), tensio-active agent are that the magnetic bead of 5%Tween-20 and 0.5 μ g is as solid phase carrier, and contain proteolytic enzyme and 6M urea, the EtOH that cleaning buffer solution is 70%.Can isolate approximately 2 μ g DNA, purity (A from the human whole blood of 200 μ l
260/ A
280) be about 1.3～1.6.
2. the separate nucleic acid of whole blood and blood plasma
The separation agent using in this embodiment contain 10% PEG, salt for 6M guanidine hydrochloride, tensio-active agent be 10%Tween-20, and the urea of magnetic solid phase carrier, proteolytic enzyme and the 6M of 0.5 μ g.
The separation agent of the present invention of getting 720 μ l directly mixes with the blood plasma after human whole blood or the centrifugation of 200 μ l, at 56 DEG C, react after 10～15 minutes, separate nucleic acid can be bonded on magnetic solid phase carrier, remove after the aqueous solution, washed with cleaning buffer solution, finally add 200 μ l aseptic deionized waters to rush the nucleic acid that dike after 10 minutes puts forward magnetic solid phase carrier upper punch and analyze.The EtOH that the cleaning buffer solution that this embodiment uses is 70%.
In the control group of embodiment, with the MegaZorb of commercially available Promega company
reagent cover group, as positive control group, represents traditional separation method, comprises that dissolving (lysis), extraction, precipitation, cleaning, punching 5 steps such as put forward.Separated DNA is analyzed, and its result as shown in Table 2.Result shows the plasma sample of separation agent of the present invention in whole blood sample or the centrifugal pre-treatment of process, all can be purified into DNA approximately 5 μ g, purity (A by 200 μ l samples
260/ A
280) being about 1.89, separation agent of the present invention can be directly used in complex samples purification of nucleic acid, does not need centrifugal Sample pretreatment.
The effect that table two, DNA separate
In another embodiment, use identical separation agent, and with the MegaZorb of commercially available Promega company
reagent cover group is as positive control group.Each experiment repeats 3 times.
From table three result, separation agent of the present invention and method can be effectively directly by DNA by separating in whole blood, and its separating effect and commercially available prod are suitable, the separable DNA of going out of 200 μ l whole blood sample measures approximately 5 μ g, this has higher purity with the DNA that separation agent of the present invention was separated, and repetitive error is less, purity (A
260/ A
280) being about 1.9 ± 0.051, the sample of nucleic acid of purifying can be applicable to polymeric enzyme reaction analysis (Polymerase Chain Reaction, PCR).
The effect that table three, DNA separate
In another embodiment, separation agent of the present invention can at room temperature react, can be effectively directly by DNA by separating in whole blood, and the separable DNA of going out of 200 μ l whole blood sample measures approximately 4 μ g, purity (A
260/ A
280) be about 1.7.
3. the separate nucleic acid of cell
The separation agent using in this embodiment contains 10% PEG, salt for 6M guanidine hydrochloride (Gu-HCl), tensio-active agent are the magnetic solid phase carrier of 10%Tween-20 and 0.5 μ g, and contains proteolytic enzyme and 6M urea.
The separation agent of the present invention of getting 720 μ l directly mixes with the CT26 mice colorectal cancer cell strain of 200 μ l, and total cellular score is about 8 × 10
5cells/ml, at 56 DEG C, reacts after 10 minutes, separate nucleic acid can be bonded on magnetic solid phase carrier, remove after the aqueous solution, be washed with cleaning buffer solution, finally add the aseptic deionized water of 200 μ l to rush dike after 10 minutes, analyze by the nucleic acid putting forward that rushes on magnetic solid phase carrier.The EtOH that the cleaning buffer solution that this embodiment uses is 70%.Its result as shown in Table 4.In this embodiment, with the MegaZorb of commercially available Promega company
reagent cover group is as positive control group.
From result, separation agent of the present invention and method can be effectively directly by DNA by separating in cell, approximately can be by 10
5cell purification goes out the DNA of approximately 7 μ g, purity (A
260/ A
280) higher than 1.8.
The effect that table four, DNA separate
The above-mentioned sample of nucleic acid being extracted by cell is directly carried out to the electrophoretic analysis of DNA agar-agar, and its result as shown in Figure 1.Reagent of the present invention and MegaZorb
all show the DNA bright band (swimming lane 1: by MegaZorb of high molecular
the nucleic acid extracting; Swimming lane 2～3: the nucleic acid being extracted by reagent of the present invention; Swimming lane 4:1Kb DNA marker).By the directly DNA in isolated cell of the known separation agent of the present invention of electrophoresis result and method, and its DNA isolation has high yield and high purity.
Although the present invention discloses as above with preferred embodiment, so it is not in order to limit the present invention, any those of ordinary skill in the art, without departing from the spirit and scope of the present invention, when doing a little change and retouching.
Claims (26)
1. a method for isolating nucleic acid, the method comprises:
After the biological specimen that one separation agent and is contained to nucleic acid mixes, described separation agent can make nucleic acid in biological specimen, separate and be bonded on the solid phase carrier being contained in described separation agent, it is characterized in that:
Ethanol, a guanidine hydrochloride, a tensio-active agent, proteolytic enzyme, urea and described solid phase carrier that described separation agent is 40～60% by polyoxyethylene glycol and/or the weight concentration of 1～40wt% are formed, and it makes nucleic acid be bonded to described solid phase carrier by the solubleness that changes nucleic acid.
2. the method for isolating nucleic acid as claimed in claim 1, the weight concentration of wherein said polyoxyethylene glycol is 5～20%.
3. the method for isolating nucleic acid as claimed in claim 1, wherein said solid phase carrier comprises metal, metal oxide, silicide or polymkeric substance.
4. the method for isolating nucleic acid as claimed in claim 1, wherein said solid phase carrier comprises magnetic substance.
5. the method for isolating nucleic acid as claimed in claim 1, wherein said tensio-active agent comprises Tween-20, Triton X-100, NP40, Brij35 or CHAPS.
6. the method for isolating nucleic acid as claimed in claim 1, the hybrid reaction temperature of wherein said reagent and biological specimen is 50 DEG C～80 DEG C.
7. the method for isolating nucleic acid as claimed in claim 1, the mixing acid base number of wherein said biological specimen and reagent is pH6～9.
8. the method for isolating nucleic acid as claimed in claim 1, wherein said biological specimen includes one or more nucleic acid.
9. the method for isolating nucleic acid as claimed in claim 1, wherein said biological specimen includes cellular material.
10. the method for isolating nucleic acid as claimed in claim 1, wherein said biological specimen comprises prokaryotic organism, fungi, virus, cell, blood, amniotic fluid, cerebrospinal fluid, or from tissue and the tissue juice thereof of skin, muscle, conjunctiva, placenta or intestines and stomach.
The method of 11. isolating nucleic acids as claimed in claim 1, the method is also included in after described biological specimen and reagent mix, carries out a wash procedure.
The method of 12. isolating nucleic acids as claimed in claim 11, the method is also included in after described wash procedure, carries out a punching and puies forward program.
The reagent of 13. 1 kinds of isolating nucleic acids, ethanol, a guanidine hydrochloride, a tensio-active agent, proteolytic enzyme, urea and a solid phase carrier that this reagent is 40～60% by polyoxyethylene glycol and/or the weight concentration of 1～40wt% are formed.
The reagent of 14. isolating nucleic acids as claimed in claim 13, the weight concentration of wherein said polyoxyethylene glycol is 5～20wt%.
The reagent of 15. isolating nucleic acids as claimed in claim 13, wherein said tensio-active agent comprises Tween-20, Triton X-100, NP40, Brij35 or CHAPS.
The reagent of 16. isolating nucleic acids as claimed in claim 13, wherein said solid phase carrier comprises metal, metal oxide, silicide or polymkeric substance.
The reagent of 17. isolating nucleic acids as claimed in claim 13, wherein said solid phase carrier comprises magnetic substance.
The cover group of 18. 1 kinds of isolating nucleic acids, this cover group comprises:
One reagent, ethanol, a guanidine hydrochloride, a tensio-active agent, proteolytic enzyme, urea and a solid phase carrier that its polyoxyethylene glycol by 1～40wt% and/or weight concentration are 40～60% are formed; And
One container.
The cover group of 19. isolating nucleic acids as claimed in claim 18, the weight concentration of wherein said polyoxyethylene glycol is 5～20wt%.
The cover group of 20. isolating nucleic acids as claimed in claim 18, wherein said tensio-active agent comprises Tween-20, Triton X-100, NP40, Brij35 or CHAPS.
The cover group of 21. isolating nucleic acids as claimed in claim 18, wherein said solid phase carrier comprises metal, metal oxide, silicide or polymkeric substance.
The cover group of 22. isolating nucleic acids as claimed in claim 18, wherein said solid phase carrier comprises magnetic substance.
The cover group of 23. isolating nucleic acids as claimed in claim 18, wherein this cover group also comprises that a cleaning buffer solution and/or a punching carry damping fluid.
The cover group of 24. isolating nucleic acids as claimed in claim 23, wherein said cleaning buffer solution can be removed salt and protein.
The cover group of 25. isolating nucleic acids as claimed in claim 23, damping fluid is carried in wherein said punching can be by nucleic acid by rinsing on solid phase carrier.
The cover group of 26. isolating nucleic acids as claimed in claim 18, wherein said container is for splendid attire separation agent.
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| CN103212170B (en) * | 2013-04-03 | 2015-07-22 | 哈尔滨工程大学 | Forest fire fighting bomb launching device |
| CN107058290A (en) * | 2017-04-28 | 2017-08-18 | 北京金豪制药股份有限公司 | A kind of method for extracting nucleic acid and its extracts reagent |
| CN111534511A (en) * | 2020-06-03 | 2020-08-14 | 沈阳德宇生物科技有限公司 | Reaction liquid for nucleic acid purification and recovery, nucleic acid recovery kit and application thereof |
| CN112321870B (en) * | 2020-11-06 | 2022-11-18 | 浙江工商大学 | Microneedle patch and preparation method and application thereof |
| WO2022135420A1 (en) * | 2020-12-23 | 2022-06-30 | 四川安可瑞新材料技术有限公司 | Preparation method for magnetic microspheres for nucleic acid extraction, prepared product and use thereof |
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