CN105754991A - RNA protecting method based on magnetic graphene nano-composite - Google Patents

RNA protecting method based on magnetic graphene nano-composite Download PDF

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CN105754991A
CN105754991A CN201610186951.1A CN201610186951A CN105754991A CN 105754991 A CN105754991 A CN 105754991A CN 201610186951 A CN201610186951 A CN 201610186951A CN 105754991 A CN105754991 A CN 105754991A
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rna
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graphene nano
magnetic graphene
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邢达
廖玉辉
周小明
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South China Normal University
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Abstract

The invention discloses an RNA protecting method based on a magnetic graphene nano-composite.According to the RNA protecting method, on the basis of a graphene oxide RNA protecting function, the magnetic graphene nano-composite having a magnetic function is synthesized; the magnetic graphene nano-composite adsorbs RNA, so that the free energy of RNA is reduced, RNA is fixed to the rigid plane of the magnetic graphene nano-composite, the purpose of effectively stopping enzymatic degradation of RNA enzyme to RNA is achieved, and finally the purpose of efficiently protecting RNA is achieved.Due to the facts that the magnetic graphene nano-composite can make a rapid response under the action of a magnetic field and is gathered to one end of the magnetic field, the RNA protecting method has the advantage of being high in protecting efficiency, separation is easy, and the RNA protecting method has important meaning for simplification of experiment steps and development of a deriving technology, and further has the advantages of being easy to operate, long in protecting time and high in capacity of resisting to RNA enzyme, and storage is easy.

Description

RNA guard method based on magnetic graphene nano-complex
Technical field
The invention belongs to RNA nanometer of resist technology field, particularly to a kind of RNA guard method based on magnetic graphene nano-complex.
Background technology
Ribonucleic acid (RNA), as important carrier of genetic information, is widely present in cell, virus and viroid, is mainly made up of mRNA, rRNA, tRNA, microRNA, sRNA, telomerase RNA.RNA in vivo and performs complicated biological function in cell: the hereditary information on DNA accurately inerrably is transcribed by mRNA, to convey hereditary information；TRNA has specific recognition, transhipment aminoacid functional；RRNA is ribosomal important composition composition；MicroRNA has adjusting function protein expression vector, the transport of aminoacid orientation, metabolic regulation etc..As can be seen here, RNA is significant for maintaining of vital movement.Explore and physiological significance research to realize RNA function, it usually needs RNA is easily separated purification, to obtaining quantitative data, as the processes such as the mensuration of specific rna expression amount, the calculating of rna expression spectrum, RNA order-checking are required for separating purifying RNA.But, once RNA leaves Cell Homeostasis and arises that serious degraded.Its readily degradable makes the preservation of RNA face huge actual challenges.At present, the protection of in vitro RNA relies primarily on RNase inhibitor, by the method stored and reaction system artificially adds RNase inhibitor, reducing and suppress the purpose of RNase activity.This type of method can only suppress or reduce but can not fundamentally block RNase activity.Plus problems such as high cost, transport difficult, perishable, active instability, the scope of application of RNase inhibitor is greatly limited.Therefore, Development of Novel, RNA guard method stable, dependable performance are significant for the protection of RNA.
In recent years, along with the high speed development of nano material, functional nanomaterials is widely used to the every field of life sciences, and achieves significant achievement.Developing the RNA guard method based on nano material, the change for existing RNA guard method is significant.My seminar's early stage patent (201510249894.2) proposes a kind of RNA guard method based on graphene oxide, and the method utilizes the graphene oxide absorption to RNA, it is achieved efficient, long-acting RNA protection.But it exists graphene oxide and separates complicated problem, taking method that is centrifugal or that filter, process is loaded down with trivial details and efficiency is low.
Summary of the invention
In order to overcome shortcoming and the deficiency of existing RNA resist technology, it is an object of the invention to provide a kind of RNA guard method based on magnetic graphene nano-complex.The RNA guard method of the present invention is: on the basis of graphene oxide RNA defencive function, and synthesis has the Graphene (magnetic graphene nano-complex) of magnetic function, and is applied to RNA protection field.The present invention adopts in RNA solution; the method adding magnetic graphene nano-complex; by the absorption to RNA of the magnetic graphene nano-complex; thus RNA being fixed on the rigid plane of magnetic graphene nano-complex; reach the effectively prevention RNase purpose to the enzymatic degradation of RNA, finally realize the purpose of efficient RNA protection.
The purpose of the present invention is achieved through the following technical solutions:
Based on the RNA guard method of magnetic graphene nano-complex, comprise the steps:
A, employing graphene oxide, as synthetic material, are utilized the carboxylic group of surface of graphene oxide as connection site, and are connected with amino magnetic bead by amido link, obtain magnetic graphene nano-complex；
B, the magnetic graphene nano-complex obtained by step A are sufficiently mixed with RNA sample or RNase, and stand, and obtain magnetic graphene-RNA mixed liquor or magnetic graphene-RNase mixed liquor；Make magnetic graphene nano-complex fully adsorb RNA, reach the purpose of protection RNA.
The mass ratio of described magnetic graphene nano-complex and RNA sample is preferably 1:(1～10)；
The ratio of described magnetic graphene nano-complex and RNase is preferably 1 μ g:(1～40U).
The time of the standing described in step B is preferably 10～30 minutes.
Described magnetic graphene-RNA mixed liquor, places 4 days at normal temperatures, and RNA is substantially without degraded；
In described magnetic graphene-RNase mixed liquor, RNase activity is substantially reduced, thus reducing the RNase degradation capability to RNA.
Based on the RNA guard method of magnetic graphene nano-complex, comprise the steps:
I. the checking of magnetic graphene nano-complex RNA protected effect: magnetic graphene nano-complex and RNA sample are sufficiently mixed; and stand 10～30 minutes; obtain magnetic graphene-RNA mixed liquor; mixed liquor is exposed in the middle of air after 24 hours, adopts the genomic integrity of electrophoresis checking R NA；
II. magnetic graphene nano-complex RNA protective value is evaluated:
1) magnetic graphene-RNA mixed liquor is exposed in air, and at the integrity of different time supervision rna gene groups, evaluates the time period of magnetic graphene RNA protection with this；
2) different RNase Concentraton gradient is set, and the RNase of variable concentrations is acted on the RNA by magnetic graphene protection, check the ultimate value of magnetic graphene nano-complex tolerance RNase with this；
III. the utilization of magnetic graphene nano-complex RNA protection.
The described RNA guard method based on magnetic graphene nano-complex, including step in detail below:
1. the synthesis of magnetic graphene nano-complex and sign
The present invention adopts graphene oxide as synthetic material, utilizes the carboxylic group of surface of graphene oxide as connection site, and is connected with amino magnetic bead by amido link, finally realizes the synthesis of magnetic graphene nano-complex；The synthetic route of magnetic graphene nano-complex as shown in Figure 1A, mainly comprises the steps that
1) graphene oxide pretreatment
The graphene oxide (particle diameter is 1 micron) taking 10mg is dissolved in 30mL distilled water, and ultrasonic one and a half hours, make graphene oxide fully dissolve, obtain the graphene oxide solution of 0.33mg/mL, and it is standby to be stored in 4 DEG C of refrigerators.
2) graphene oxide activated carboxylic
To step 1) in gained graphene oxide solution adds 4mgN-N-Hydroxysuccinimide (NHS), and after fully mixing, add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) of 5mg, fully after mixing, stirring reaction 1 hour at 37 DEG C, and ultrasonic half an hour make the graphene oxide of activated carboxylic be again dispersed into monolayer without assemble solution.
3) amido link connects
To step 2) solution of gained adds 10mg amino magnetic bead (particle diameter is 200nm), and fully mix.After sealing, 37 DEG C of stirring reactions cross liquid, obtain the thick product of magnetic graphene nano-complex.
4) purification
Step 3) in the thick product of gained, be not connected with the graphene oxide of magnetic bead containing small part, it is necessary to be purified to remove in solution and do not have magnetic graphene oxide.The present invention adopts the method for Magneto separate, the magnetic graphene oxide of tool is made to converge at one end, magnetic field, removal does not have magnetic supernatant solution, add isopyknic PBS (concentration is 1 ×), after removing magnetic field, mixing is shaken, and then carries out Magneto separate operation again and can obtain purer magnetic graphene nano-complex for 2 times.
2. the checking of magnetic graphene nano-complex RNA protected effect
Magnetic graphene nano-complex RNA protection philosophy is such as shown in Figure 1B, C:
1. Total RNAs extraction thing 50 μ L is taken.
2. 1 μ L magnetic graphene nano-complex (1mg/mL) is taken, add step 1. described in Total RNAs extraction thing, and the water (without RNase) adding 49 μ L complements to 100 μ L, and concussion mixes, and room temperature place 10～30 minutes, make magnetic graphene fully adsorb RNA.And it is numbered experimental group 1.
3. gained Total RNAs extraction thing 50 μ L in step 1 is separately taken.Add the water without RNase and complement to 100 μ L.Repeat 3 times, and numbering experimental group 2,3,4.Wherein experimental group 2 and 4 is left intact respectively as the matched group of experimental group 1 and 3, puts into-80 refrigerators stand-by.
4. by experimental group 1 and experimental group 3, uncapping and be exposed in air, room temperature is placed 24 hours.
5. take the experimental group 1 and 3 after step 4. middle process, carry out agarose gel electrophoresis experiment (voltage: 120V, 45min).And the intact degree of analytical electrophoresis band, evaluation RNA.Experimental group 2 and 4 is taken out from-80 refrigerators, and and experimental group 1 and 3 comparing calculation degradation rate.
3. magnetic graphene nano-complex RNA protective value is evaluated
1) the magnetic graphene nano-complex RNA guard time limit
1. take 750 μ L Total RNAs extraction things, and add 15 μ L magnetic graphene nano-complex stock solution (1mg/mL), fully after mixing, equivalent subpackage to the centrifuge tube of 15 200 μ L, often pipe 51 μ L.
2., respectively in above-mentioned centrifuge tube, the 49 μ L pure water without RNase is added.Fully mixing is standby.
3. above-mentioned 15 groups of samples are exposed in the middle of air, and room temperature is placed.
4. every 24 hours, take a pipe sample, and evaluate the integrated degree of Total RNAs extraction thing in each experimental group with agarose gel electrophoresis.The monitoring process of 15 days is completed with this.
5. collect experimental result, and the intact degree and variation tendency according to 15 sample Total RNAs extraction things goes to evaluate the limit of the RNA guard time that magnetic graphene nano-complex provides.
2) the magnetic graphene nano-complex tolerance RNase limit
1. take 12 μ L magnetic graphene nano-complex (initial concentration 1mg/mL), and be all divided in 200 μ L centrifuge tubes, often organize 1 μ L.
2. RNase original liquid concentration gradient is set, from 10U to 120U, one experimental group is set every 10U, obtain the RNase solution of gradient dilution.
3. to step 1. described in magnetic graphene nano-complex in add (100ng/ μ L) Total RNAs extraction thing of equivalent (1 μ L), and add the RNase solution of gradient dilution therewith.Fully mixing, obtains 12 groups of magnetic graphene-total serum IgE-RNase mixed liquors, standby.
4. the mixed liquor 37 DEG C of step 3. middle gained is hatched 30min.
5. taking out step 4. middle reactant liquor 8 μ L, add RNA fluorescent dye (SYBRGold, 10 ×) 2 μ L, mix and dye 10min.
6. previously prepared good agarose gel is taken, for the electrophoretic analysis (voltage: 120V, 45min) of step 4. described mixed liquor.Gel imaging, and observed and recorded experimental result.According to the integrated degree of RNA in electrophoresis result and variation tendency, it is judged that the limit capacity of magnetic graphene nano-complex tolerance RNase.
Principles of the invention: the present invention utilizes the absorption to RNA of the magnetic graphene nano-complex, thus reducing its free energy, finally realizes the purpose of RNA protection.Owing to magnetic graphene nano-complex is under the action of a magnetic field, it is possible to make quick response, and be gathered in one end, magnetic field.Therefore, the RNA guard method based on magnetic graphene nano-complex not only has the feature that protective efficacy is high, and is easily isolated, and the development for simplifying experimental procedure and deriving technology is significant.
The present invention, relative to prior art, has such advantages as and effect:
(1) RNA protective efficacy is high, and RNA can be provided effectively protection by a small amount of magnetic graphene nano-complex.
(2) being easily isolated, magnetic graphene nano-complex under the influence of a magnetic field, can make quick response, and separation process is simple.
(3) simple to operate, guard time is long, exposes in aerial situation at RNA, can provide the protection up to four days for RNA.
(4) ability of anti-RNase is strong.
(5) easily stored, it is positioned over 4 DEG C of refrigerators.
(6) magnetic graphene good dispersion, magnetic response time are short.
Accompanying drawing explanation
Fig. 1 is the RNA protection mechanism schematic diagram of magnetic graphene nano-complex of the present invention；Wherein, Figure 1A. magnetic graphene nano-complex synthetic route；Figure 1B, C. magnetic graphene nano-complex RNA protection philosophy.
Fig. 2 is the comparing result of magnetic bead (MB), graphene oxide (GO), magnetic graphene nano-complex；Wherein, Fig. 2 A. magnetic bead (MB), graphene oxide (GO), magnetic graphene nano-complex (MGO) comparison diagram；Fig. 2 B. graphene oxide atomic force microscope phenogram；Fig. 2 C. magnetic graphene nano-complex atomic force phenogram；Fig. 2 D. nanometer particle size phenogram；Fig. 2 E.Zeta current potential phenogram.
Fig. 3 is the result figure of magnetic graphene nano-complex magnetic response assemble index.
Fig. 4 is magnetic bead and magnetic graphene nano-complex dispersibility comparing result figure.
Fig. 5 is the result figure of magnetic graphene nano-complex RNA protection；Wherein, Fig. 5 A. magnetic graphene nano-complex RNA protects electrophoretogram and degradation rate；Fig. 5 B. magnetic graphene nano-complex RNA protects electrophoresis gray analysis.
The time period that Fig. 6 is embodiment 3 magnetic graphene nano-complex RNA protective effect is probed into；Wherein, Fig. 6 A. magnetic graphene nano-complex RNA guard time limit；Fig. 6 B. magnetic graphene nano-complex tolerance RNase limit.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
The synthesis of embodiment 1 magnetic graphene nano-complex and sign
The present invention adopts graphene oxide as synthetic material, utilizes the carboxylic group of surface of graphene oxide as connection site, and is coated magnetic bead by amido link with amino and is connected, and finally realizes the synthesis of magnetic graphene nano-complex.The synthetic route of magnetic graphene nano-complex as shown in Figure 1A, mainly comprises the steps that
1) graphene oxide pretreatment
The graphene oxide (particle diameter is 1 micron) taking 10mg is dissolved in 30mL distilled water, and ultrasonic one and a half hours, make graphene oxide fully dissolve, obtain the graphene oxide solution of 0.33mg/mL, and it is standby to be stored in 4 DEG C of refrigerators.
2) graphene oxide activated carboxylic
To step 1) in gained graphene oxide solution adds 4mgN-N-Hydroxysuccinimide (NHS), and after fully mixing, add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) of 5mg, fully after mixing, stirring reaction 1 hour at 37 DEG C, and ultrasonic half an hour make the graphene oxide of activated carboxylic be again dispersed into monolayer without assemble solution.
3) amido link connects
To step 2) solution of gained adds 10mg amino magnetic bead (particle diameter is 200nm), and fully mix.After sealing, 37 DEG C of stirring reactions cross liquid, obtain the thick product of magnetic graphene nano-complex.
4) purification
Step 3) in the thick product of gained, be not connected with the Graphene of magnetic bead containing small part, it is necessary to be purified to remove in solution and do not have magnetic graphene oxide.The present invention adopts the method for Magneto separate, the magnetic graphene oxide of tool is made to converge at one end, magnetic field, removal does not have magnetic supernatant solution, add isopyknic PBS (concentration is 1 ×), after removing magnetic field, mixing is shaken, and then carries out Magneto separate operation again and can obtain purer magnetic graphene nano-complex for 2 times.
By Fig. 2 A it can be seen that relative to graphene oxide, magnetic graphene nano-complex in character with deeper of black.When there being magnetic field to get involved, magnetic graphene converges at the side near magnetic field, and solution is become colourless by black.In addition, magnetic graphene nano-complex has been done atomic force and has characterized experiment by us, experimental result is such as shown in Fig. 2 B and C: graphene oxide presents the pattern of lamellar, after graphene oxide and magnetic bead connect, laminated structure occurs in that spherical projection, thus demonstrates the feasibility of magnetic graphene nano-complex composition principle of the present invention.Nanometer particle size contrast experiment (Fig. 2 D) shows, magnetic graphene nano-complex has bigger particle diameter (about 2 microns) relative to independent magnetic bead and graphene oxide.Zeta potential characterizes experiment display graphene oxide in stronger negative charge, and relative to graphene oxide, magnetic graphene nano-complex presents more weak negative charge, and this has neutralized the negative charge of graphene oxide self owing to the positive charge of amino magnetic bead self.Thus, the feasibility of the synthetic route of magnetic graphene nano-complex obtains sufficient proof.
In order to verify the magnetic response time of magnetic graphene nano-complex, at different time points (2 minutes, 4 minutes, 6 minutes, 8 minutes, 10 minutes), observe magnetic graphene nano-complex assemble index under the action of a magnetic field.Experimental result is as it is shown on figure 3, magnetic graphene nano-complex under the influence of a magnetic field, is gathered in magnetic field end gradually, and when 4 minutes, major part magnetic graphene was gathered in magnetic field end.Additionally, it has been found that magnetic graphene is relative to independent amino magnetic bead, dispersibility is better (see Fig. 4).Amino magnetic bead when standing 10 minutes after mostly fall to flat, and magnetic graphene nano-complex remains stable in 25min and does not settle.
Embodiment 2 magnetic graphene nano-complex RNA protective value is verified
Magnetic graphene nano-complex RNA protection philosophy is such as shown in Figure 1B, C:
1) checking magnetic graphene protects RNA ability in atmosphere
1. Total RNAs extraction thing 50 μ L (100ng/ μ L) is taken.
2. 1 μ L magnetic graphene nano-complex (1mg/mL) is taken, add step 1. described in Total RNAs extraction thing, and the water (without RNase) adding 49 μ L complements to 100 μ L, and concussion mixes, and room temperature place 10～30 minutes, make magnetic graphene fully adsorb RNA.And it is numbered experimental group 1.
3. gained Total RNAs extraction thing 50 μ L in step 1 is separately taken.Add the water without RNase and complement to 100 μ L.Repeat 3 times, and numbering experimental group 2,3,4.Wherein experimental group 2 and 4 is left intact respectively as the matched group of experimental group 1 and 3, puts into-80 refrigerators stand-by.
4. by experimental group 1 and experimental group 3, uncapping and be exposed in air, room temperature is placed 24 hours.
5. take the experimental group 1 and 3 after step 4. middle process, carry out agarose gel electrophoresis experiment (voltage: 120V, 45min).And the intact degree of analytical electrophoresis band, evaluation RNA.Experimental group 2 and 4 is taken out from-80 refrigerators, and and experimental group 1 and 3 comparing calculation degradation rate.
Experimental result is as it is shown in figure 5, experimental group 3 occurs in that obvious degraded, and experimental group 1 (RNA by magnetic graphene protection) still remains stable for, and thus demonstrates the RNA protective capability of magnetic graphene nano-complex.Meanwhile, degradation rate data show, under the protection of magnetic graphene nano-complex, the degradation rate of RNA substantially reduces.
Embodiment 3 magnetic graphene nano-complex RNA protective value is evaluated
1) the magnetic graphene nano-complex RNA guard time limit
1. take 750 μ L Total RNAs extraction thing 100ng/ μ L, and add 15 μ L magnetic graphene nano-complex (1mg/mL), fully after mixing, equivalent subpackage to the centrifuge tube of 15 200 μ L, often pipe 51 μ L.
2., respectively in above-mentioned centrifuge tube, the 49 μ L pure water without RNase is added.Fully mixing is standby.
3. above-mentioned 15 groups of samples are exposed in the middle of air, and room temperature is placed.
4. every 24 hours, take a pipe sample, and evaluate the integrated degree of Total RNAs extraction thing in each experimental group with agarose gel electrophoresis.The monitoring process of 15 days is completed with this.
5. collect experimental result, and the intact degree and variation tendency according to 15 sample Total RNAs extraction things goes to evaluate the limit of the RNA guard time that magnetic graphene nano-complex provides.
As shown in Figure 6A, magnetic graphene nano-complex can provide the protection up to 4 days for RNA to experimental result.And do not add the RNA of magnetic graphene nano-complex, occur in that serious degraded.
2) the magnetic graphene nano-complex tolerance RNase limit
1. take 12 μ L magnetic graphene nano-complex (initial concentration 1mg/mL), and be all divided in 200 μ L centrifuge tubes, often organize 1 μ L.
2. RNase I original liquid concentration gradient is set, from 10U to 120U, one experimental group is set every 10U, obtain the RNase solution of gradient dilution.
3. to step 1. described in magnetic graphene nano-complex in add (100ng/ μ L) Total RNAs extraction thing of equivalent (1 μ L), and add the RNase solution of gradient dilution therewith.Fully mixing, obtains 12 groups of magnetic graphene-total serum IgE-RNase mixed liquors standby.
4. the mixed liquor 37 DEG C of step 3. middle gained is hatched 30min.
5. taking out step 4. middle reactant liquor 8 μ L, add RNA fluorescent dye (SYBRGold, 10 ×) 2 μ L, mix and dye 10min.
6. take step 5. in previously prepared good agarose gel, for the electrophoretic analysis (voltage: 120V, 45min) of step 4. described mixed liquor.Gel imaging, and observed and recorded experimental result.According to the integrated degree of RNA in electrophoresis result and variation tendency, it is judged that the limit capacity of magnetic graphene nano-complex tolerance RNase.
By Fig. 6 B it can be seen that graphene oxide can be effectively protected for RNA offer.The degraded of 40URNA enzyme can be resisted.
Above-described embodiment is the present invention preferably embodiment; but embodiments of the present invention are also not restricted to the described embodiments; the change made under other any spirit without departing from the present invention and principle, modification, replacement, combination, simplification; all should be the substitute mode of equivalence, be included within protection scope of the present invention.

Claims (5)

1. based on the RNA guard method of magnetic graphene nano-complex, it is characterised in that comprise the steps:
A, employing graphene oxide, as synthetic material, are utilized the carboxylic group of surface of graphene oxide as connection site, and are connected with amino magnetic bead by amido link, obtain magnetic graphene nano-complex；
B, the magnetic graphene nano-complex obtained by step A are sufficiently mixed with RNA sample or RNase, and stand, and obtain magnetic graphene-RNA mixed liquor or magnetic graphene-RNase mixed liquor.
2. the RNA guard method based on magnetic graphene nano-complex according to claim 1, it is characterised in that:
The mass ratio of described magnetic graphene nano-complex and RNA sample is 1:(1～10).
3. the RNA guard method based on magnetic graphene nano-complex according to claim 1, it is characterised in that:
The ratio of described magnetic graphene nano-complex and RNase is 1 μ g:(1～40U).
4. the RNA guard method based on magnetic graphene nano-complex according to claim 1, it is characterised in that:
The time of the standing described in step B is 10～30 minutes.
5. the RNA guard method based on magnetic graphene nano-complex according to claim 1, it is characterised in that:
Described magnetic graphene-RNA mixed liquor, places 4 days at normal temperatures, and RNA is substantially without degraded.
CN201610186951.1A 2016-03-29 2016-03-29 RNA protecting method based on magnetic graphene nano-composite Pending CN105754991A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107236730A (en) * 2017-07-14 2017-10-10 博奥生物集团有限公司 A kind of SPE material and its application in the enrichment and detection of nucleic acid
CN109956469A (en) * 2017-12-26 2019-07-02 中国科学院苏州纳米技术与纳米仿生研究所 The enzymatic degradation method and its application of three-dimensional grapheme

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Publication number Priority date Publication date Assignee Title
CN104805076A (en) * 2015-05-15 2015-07-29 华南师范大学 Novel application of graphene oxide as RNA (ribonucleic acid) protection reagents

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Publication number Priority date Publication date Assignee Title
CN104805076A (en) * 2015-05-15 2015-07-29 华南师范大学 Novel application of graphene oxide as RNA (ribonucleic acid) protection reagents

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夏前芳: "新型石墨烯传感材料的构建及应用研究", 《中国优秀硕士学位论文全文数据库 信息科技辑》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107236730A (en) * 2017-07-14 2017-10-10 博奥生物集团有限公司 A kind of SPE material and its application in the enrichment and detection of nucleic acid
CN109956469A (en) * 2017-12-26 2019-07-02 中国科学院苏州纳米技术与纳米仿生研究所 The enzymatic degradation method and its application of three-dimensional grapheme

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