CN106501343A - A kind of method that heavy metal ion is detected based on polymerase isothermal amplification technique - Google Patents
A kind of method that heavy metal ion is detected based on polymerase isothermal amplification technique Download PDFInfo
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- CN106501343A CN106501343A CN201610905817.2A CN201610905817A CN106501343A CN 106501343 A CN106501343 A CN 106501343A CN 201610905817 A CN201610905817 A CN 201610905817A CN 106501343 A CN106501343 A CN 106501343A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
Abstract
The present invention relates to a kind of method for detecting heavy metal nickel ion based on magnetic core-shell nanoparticle and polymerase isothermal amplification signal amplification technique, belongs to analytical chemistry or environmental monitoring technology field.Using self-assembling technique, DNA S1 is fixed to Fe by Au S keys3O4@Au core-shell magnetic nanoparticles surface.Nickel ion（Ni2+）In the presence of, nucleic acid chains S1 are in enzyme site by Ni2+Cutting, the number of base disappearance at 5 ' ends of nucleic acid chains S1 cause its 3 ' end nucleic acid chains to become single-stranded, and single-stranded S2 generations hybridization is combined with the DNA of another complementary seriess then.After polymerase and substrate dNTP is added, under the induction progradation of S2 S1 is promoted to open and isothermal duplication iodine occurs, methylene blue MB is embedded into double-stranded DNA space finally, the complex of formation realizes the amplification of electrochemical response signal on golden carbon electrode surface by magnetite gathering.Realized to Ni in solution according to the enhancing of electrochemical signals2+Measure, the method have high sensitivity, high selectivity, simple, quick the features such as.
Description
Technical field
The invention belongs to analytical chemistry or environmental monitoring technology field, and in particular to a kind of based on magnetic nano-particle and poly-
Synthase isothermal amplification signal amplification technique detects the electrochemical method of heavy metal nickel ion.
Background technology
Nickel is to maintain trace element necessary to body metabolism process, and its biological action is extremely wide, but nickel and
Its compound is common environmental pollution factors again.Nickel is widely used in the row such as plating, automobile, electrolysis, pesticide, medicine, paint
Industry.Nickel can enter human body by air suction and the number of ways such as diet, can cause when savings is excessive in vivo nasal cavity, lung and
A series of diseases such as heart, or even induction cancer.Therefore, nickel is to need one of heavy metal of monitoring in the media such as environment, food.
At present, the conventional assay method of nickel has spectrophotography, atomic spectroscopy, inductively coupled plasma mass spectrometry and fluorescence method
Deng.These methods not only need the instrument of high precision, and operating technology is had high demands, and detection cycle length, testing cost are high.Cause
This, needs development simple, quick, and can achieve new method and the new technique of trace detection nickel.
Nucleic acid isothermal amplification technology is a kind of brand-new Progress of Nucleic Acid Amplification Technologies that developed recently gets up, its course of reaction
All the time maintain at a constant temperature, by adding the enzyme and respective specific primer of different activities（Or be not added with）Quick to reach
The purpose of nucleic acid amplification.Compared with Standard PCR technology, it is not necessary to the thermal denaturation of template, temperature cycles, electrophoresis and ultraviolet visualization etc.
Process.Requirement of the nucleic acid isothermal amplification technology to instrument is greatly simplified, and the response time greatly shortens, with simple, quick, special
The characteristics of property is strong, has been widely used in the detection of all kinds of nucleic acid, virus and microorganism.1994, by in-vitro screening skill
Art obtains DNAzyme（DNAzyme）And its discovery of catalytic performance, promote the fast development of nucleic acid isothermal amplification technology.De-
Oxygen ribozyme is the single stranded DNA that the class obtained using directed evolution technology has high-efficiency catalytic activity and structure recognition ability
Molecule.Similar to RNA, DNAzyme can form complicated and diversified foldable structure and show catalysiss, can be with catalytic dna
Or RNA cutting, DNA connection, DNA hydrolysis etc. multiclass reaction.Compared with traditional protease, DNAzyme has stability high, phase
Little to molecular mass, chemically reactive synthesis, modification and replicate, by the advantage such as the such environmental effects such as acidity are little.DNAzyme is very
Easily carry out with nucleic acid as recognition mode or the biosensor of signal representation pattern builds.Additionally, DNAzyme have right
DNA is attached the catalytic capability with fracture, may apply to gene regulation and treatment.
Magnetic nano-particle is a kind of novel nano-material developed in recent years, as magnetic nano-particle has spy
Different magnetic conductance tropism, superparamagnetism, easy cleaning, easily separated, and surface can connect the characteristics such as biochemical activity functional group so as to
Application in fields such as foranalysis of nucleic acids, clinical diagnosises, targeted drug, cell separation and enzyme immobilizations has obtained extensive development.
These characteristics of magnetic nano-particle can significantly increase the sensitivity of biosensor detection, shorten the biochemical reaction time,
Detection flux is improved, is that the application of field of biosensors opens wide prospect.
The present invention is based on magnetic nano-particle and polymerase isothermal amplification as signal amplification technique, using Ni2+Right
The RNA locus specificities cutting of DNA enzymatic and electrochemical method, realize heavy metal Ni2+Ultra trace detection.In order to increase analysis
There is the long chain DNA complex of polymerase isothermal amplification formation using golden magnetic in speed and stability, magnetic nanometer surface
Electrode adsorption is fixed, and the method has simple, quick, sensitivity high.
Content of the invention
An object of the present invention is to design not homotactic DNA, builds a kind of quick, sensitive electrochemical sensor.
The second object of the present invention is to provide one kind and amplifies skill based on magnetic nano-particle and polymerase isothermal amplification
Art combination builds the preparation method of electrochemical sensor.
The third object of the present invention is highly sensitive, the quick inspection that the electrochemical sensor that will be built is used for heavy metal ion
Survey.
The concrete technical scheme of the present invention is as follows：
1. a kind of electrochemical based on magnetic nano-particle and polymerase isothermal amplification amplifying technique combination structure heavy metal ion
Learn sensor
Fe is prepared first3O4DNA nucleic acid chains S1 are fixed to Fe by Au-S keys by@Au core-shell structure copolymer magnetic nano-particles3O4@Au tables
Face.Ni2+In the presence of, nucleic acid chains S1 are in rA sites by Ni2+Two sections are cut into, cause its 3 ' end nucleic acid chains to become single-stranded, the 3 ' of S1
There is hybridization and combine in the single-stranded S2 single-stranded with the DNA of another complementary seriess in end.After polymerase and substrate dNTP is added, S2's
S1 is promoted to open and isothermal duplication iodine occurs under induction progradation, finally by methylene blue（MB）It is embedded into double-strand
DNA spaces.The long chain DNA complex for being formed realizes putting for electrochemical response signal by magnetite gathering on golden carbon electrode surface
Greatly.Realized to Ni in standard solution or sample solution according to the enhancing of electrochemical signals2+The quantitative determination of concentration.
2. a kind of combination based on magnetic nano-particle and polymerase isothermal amplification amplifying technique builds heavy metal ion
The preparation method of electrochemical sensor
（1）Fe3O4The preparation of@Au core-shell structure copolymer magnetic nano-particles
Fe is prepared initially with coprecipitation3O4, by FeCl2、FeCl3It is dissolved in ultra-pure water with dense HCl, ultrasonic deoxidation.Will be above-mentioned
Mixed solution is added drop-wise in NaOH solution, logical N under 80 DEG C of stirrings2Gas shielded, obtains Fe3O4Magnetic nano-particle.In Fe3O4/ second
3- aminopropyl triethoxysilanes are added in alcoholic solution, to Fe3O4Particle surface carries out amination.Add in above-mentioned solution
HAuCl4Aqueous solution, with sodium citrate as reducing agent, obtains Fe3O4@Au core-shell structure copolymer magnetic nano-particles.
（2）Double-stranded DNA flag F e3O4The preparation of@Au particles
By step（1）The Fe for obtaining3O4@Au magnetic nano-particles react 12 h at 37 DEG C with DNA nucleic acid chains S1, hatch, magnetic
Separate, obtain the Fe of S1 labellings3O4@Au.By Ni2+Fe with S1 labellings3O4@Au hatch 1 h at 37 DEG C, and nucleic acid chains S1 are in DNA
Enzyme site is by Ni2+Two sections are cut into, the number of base disappearance at 5 ' ends of nucleic acid chains S1 causes its 3 ' end nucleic acid chains to become single-stranded, it
5 min are heated after 95 DEG C, is caused S2 hybridize 3 H-shapeds into double-stranded DNA with the ends of S1 3 ' complementary single strand at 37 DEG C, is obtained double-strand
The Fe of DNA marker3O4@Au magnetic nano-particles.
（3）Polymerase isothermal amplification
In step（2）The Fe of the double-stranded DNA labelling for obtaining3O4Polymerase and substrate is added in@Au Tris-HCl buffer
DNTP, 2 h of hybridization at 37 DEG C, obtain long chain DNA complex.Finally, DNA complex and methylene blue（MB）Anti- at 37 DEG C
3 h are answered, MB is embedded into Fe3O4In the double-stranded DNA on@Au surfaces.
（4）Ni2+The preparation of electrochemical sensor
By working electrode（Golden carbon electrode）1 h is soaked in piranha solution, and ultra-pure water is cleaned.Then respectively with 0.3 mm and
The Al of 0.05 mm2O3Polishing powder is polished, and is cleaned by ultrasonic successively in ultra-pure water, dehydrated alcohol, ultra-pure water.Finally in 0.5 M
H2SO4In solution, it is circulated voltammetric scan 30 between -0.2 ~ 1.6 V and encloses with the speed of sweeping of 50 mV/s, activated electrode.N2Gas
Dry up standby.Step（3）The long chain DNA complex for obtaining acts on modification to the golden carbon electrode surface of activation by magnetite gathering,
Build heavy metal Ni2+Electrochemical sensor.
3. Ni2+The detection method of ion
（1）Tested with three-electrode system using electrochemical workstation, saturated calomel electrode is reference electrode, platinum electrode is
To electrode, the golden magnetoelectricity extremely working electrode of modification, in the PBS buffer solution of 10 mM pH 7.4（Comprising 20 mM NaCl and
0.1 M NaClO4）In tested；
（2）Use Differential Pulse Voltammetry（DPV）Heavy metal ion is detected that it is 0 ~ 1.2 V to arrange scanning potential range, sweeps
Speed is retouched for 0.1 V/S, the size of peak current is recorded, and is preserved data；
（3）Size and Ni according to gained DPV peak currents2+The relation of ion concentration, drawing curve；
（4）Using standard addition method, the Ni of variable concentrations is added in water sample solution2+Ion standard solution, according to working curve
Ni of the method for drafting to variable concentrations2+Ion is detected.
Compared with prior art, the present invention has advantages below and useful achievement：
（1）The present invention builds a huge sum of money using magnetic nano-particle and the combination of polymerase isothermal amplification dual signal amplifying technique
Category ion electrochemical sensor.
（2）The present invention is by Fe3O4@Au core-shell structure copolymer magnetic nano-particles are incorporated in the structure of electrochemical sensor, using gold
Carbon electrode, so that increased the stability of modified electrode.
（3）The present invention utilizes Ni2+To DNA enzymatic specificity dissection, the DNA of particular sequence is designed, so as to enhance electricity
The selectivity of pole heavy metal ion, increased the specificity of sensor.
（4）Electrochemical sensor prepared by the present invention is used for heavy metal Ni2+The detection of ion, with higher selectivity and
Capacity of resisting disturbance, range of linearity width, test limit are low, can be used for ultra trace Ni in actual water sample2+The measure of ion.
Description of the drawings
Fig. 1 is magnetic nano-particle and the polymerase isothermal amplification amplifying technique detection heavy metal using the present invention
Ni2+Schematic diagram；
Fig. 2 is the magnetic nano-particle and polymerase isothermal amplification technique detection variable concentrations Ni using the present invention2+DPV bent
Line chart；
Fig. 3 is the magnetic nano-particle and polymerase isothermal amplification technique detection Ni using the present invention2+Canonical plotting, its
Middle abscissa is Ni2+Concentration, unit is mM or M, and vertical coordinate is DPV peak currents, and unit is mA.
Specific embodiment
The present invention is further illustrated with reference to specific embodiment, but does not constitute the further restriction to inventing.
Embodiment 1 is a kind of to detect a huge sum of money based on magnetic nano-particle and the combination of polymerase isothermal amplification amplifying technique
Category Ni2+Method
The present invention is based on magnetic nano-particle and polymerase isothermal amplification amplifying technique combination detection heavy metal Ni2+Side
Method principle is as shown in Figure 1.Nucleic acid chains S1 are fixed to Fe by Au-S keys3O4@Au magnetic nano-particles surface.Ni2+In the presence of, core
Sour chain S1 is in DNA enzymatic site by Ni2+Cutting, the number of base disappearance at 5 ' ends of nucleic acid chains S1 cause its 3 ' end nucleic acid chains to become single
Chain, and then with the DNA of another complementary seriess there is hybridization combination in single-stranded S2.After polymerase and substrate dNTP is added, in S2
Induction progradation under promote S1 to open and isothermal duplication iodine occurs.MB is embedded into double-stranded DNA space, shape finally
Into long chain DNA complex the amplification of electrochemical response signal is realized on golden carbon electrode surface by magnetite gathering.According to electrification
The enhancing for learning signal is realized to Ni in standard solution or sample solution2+The quantitative determination of concentration.
2 oligonucleotide sequence of embodiment is designed
Oligonucleotide sequence designed by the present invention is by Chinese Shanghai Sangon life work biological engineering Shanghai（Share）Company limited
Synthesis, and checked by HPLC purification, lyophilizing.The oligonucleotide sequence of present invention design is as follows：
S1：5 '-ACT CAC TAT RA GGA AGA GAT GGA CGT GAG TCG ACT AGA CAC GTC CAT CTC
TGC AGT CGG GTA GTT AAA CCG ACC TTC AGA CAT AGT GAG TAG CA -3’-SH；
S2：5’- CTA CTC ACT ATG TCT -3’；
Oligonucleotide is dissolved in ultrapure aquesterilisa, -18 DEG C save backup.
3 Fe of embodiment3O4The preparation of@Au core-shell structure copolymer magnetic nano-particles
Take 1 g FeCl2×4H2O、2.6 g FeCl3×6H2O and the dense HCl of 0.425 mL are dissolved in 100 mL water, ultrasonic deoxidation.
Above-mentioned mixed liquor is added drop-wise in 125 mL, 0.75 M NaOH solutions, N2Lower 80 DEG C of stirrings are protected, Fe is obtained3O4Magnetic Nano
Particle.Prepare the Fe of 5 g/L3O425 mL of/ethanol solution, ultrasonic disperse, 0.4 mL 3- aminopropyl triethoxysilanes of Deca,
7 h are stirred under room temperature, are diluted to 1 g/L.It is separately added into 14 mL, 0.6 M HAuCl4×4H20.2 M citric acids of O and 0.3 mL
Sodium solution, supersound process under room temperature, solution color from pale yellow are changed into black, obtain Fe3O4@Au core-shell structure copolymer magnetic nano-particles.
4 oligonucleotide S1 and S2 flag F e of embodiment3O4@Au core-shell structure copolymer magnetic nano-particles
Take 1 mg Fe3O4@Au particles are scattered in the S1 solution of 1 mL, 0.6 mM, and 37 DEG C of waters bath with thermostatic control mix 12 h, PBS
Buffer solution for cleaning, is obtained the Fe of S1 labellings3O4@Au particles.Take the Fe of 60 mL S1 labellings3O4@Au and 60 mL variable concentrations
Ni2+Solution hatches 1 h, Tris-HCl buffer solution for cleaning, 95 DEG C of 5 min of heating in 37 DEG C of waters bath with thermostatic control.60 mLs 1 are taken again
MM S2 and 3 h of above-mentioned solution hybridization, is obtained the Fe of S1 and S2 double-stranded DNA labellings3O4@Au core-shell structure copolymer magnetic nano-particles.
5 polymerase isothermal amplification of embodiment
Take double-stranded DNA flag F e obtained in 60 mL embodiments 43O4@Au core-shell structure copolymer magnetic nano-particles, with 20 mM pH 7.4
Tris-HCl（Contain 60 mM KCl and 10 mM MgCl）Above-mentioned solution is diluted to 600 mL, then with 200 mM dNTP and 25
U/mL polymerases 2 h of hybridization at 37 DEG C, Tris-HCl buffer solution for cleaning are obtained long chain DNA complex modification
Fe3O4@Au particles.Finally hybridize 3 h, Tris-HCl buffer solution for cleaning at 37 DEG C with 20 mM MB again.
Prepared by 6 electrochemical sensor of embodiment
Golden carbon electrode is soaked 1 h in piranha solution, ultra-pure water is cleaned.Then respectively with 0.3 mm's and 0.05 mm
Al2O3Polishing powder is polished, and is cleaned by ultrasonic successively in ultra-pure water, dehydrated alcohol.In 0.5 M H2SO4In solution, with 50 mV/s
Speed of sweeping be circulated voltammetric scan 30 between -0.2 ~ 1.6 V and enclose, activated electrode.N2Air-blowing is done standby.By the gold that handles well
In solution obtained by carbon electrode immersion embodiment 5, golden carbon electrode surface is fixed in using magnetite gathering effect, is obtained
Ni2+Electrochemical sensor.
The detection of 7 heavy metal ion of embodiment
Tested with three-electrode system using electrochemical workstation, saturated calomel electrode is reference electrode, and platinum electrode is right
Electrode, golden magnetoelectricity extremely working electrode.Using Differential Pulse Voltammetry heavy metal Ni2+Concentration, with 10 mM pH 7.4
PBS is (containing 20 mM NaCl and 0.1 M NaClO4) be buffer, potential range be 0 ~ 1.2V, current potential amplification be 4 mV, arteries and veins
The cycle is rushed for 0.5 s.Differentiated pulse curve peak current and different Ni2+The relation curve of concentration as shown in Fig. 2 DPV peak currents with
Ni2+Concentration assumes good linear relationship in the range of 100 aM ~ 1 mM（Fig. 3）, correlation coefficientRFor 0.9962, linear side
Cheng Weii pc(mA) = 2.01logc(M)+37.56, detection is limited to 10 aM.Sensor proposed by the present invention compares which
His sensor, with the broader range of linearity and lower detection limit, anti-using magnetic nano-particle and polymerase isothermal duplication
Amplifying technique is answered to can achieve Ni2+Ultra trace detection.
8 sample determination of embodiment
Testing sample solution is replaced Ni2+Ion standard solution, using standard addition method, according to standard curve method to sample
Ni in solution2+It is measured.Measurement result shows, testing result and the Ni for adding2+Concentration of standard solution is consistent, with full
The response rate and relative standard deviation of meaning, electrochemical sensor prepared by the present invention can be used for Ni in actual sample2+Measure.
The embodiment of the present invention is illustrated above in conjunction with accompanying drawing, but the invention is not restricted to above-described embodiment, can be with
Purpose according to the innovation and creation of the present invention makes multiple changes, under all spirit and principle according to technical solution of the present invention
The change made, modification, replacement, combination, simplify, should be equivalent substitute mode, as long as meeting the goal of the invention of the present invention, only
Otherwise heavy metal ion is built based on magnetic nano-particle and the combination of polymerase isothermal amplification amplifying technique away from the present invention
The know-why of the preparation method of electrochemical sensor, ultra trace heavy metal ion electrochemical determination method and application and invention structure
Think, belong to protection scope of the present invention.
Sequence table
Raw work biological engineering（Shanghai）Limited company
DNA artificial sequences
S1：5'-ACT CAC TAT RA GGA AGA GAT GGA CGT GAG TCG ACT AGA CAC GTC CAT CTC
TGC AGT CGG GTA GTT AAA CCG ACC TTC AGA CAT AGT GAG TAG CA -3'-SH；
S2：5'-CTA CTC ACT ATG TCT-3'；
Claims (7)
1. a kind of electrochemical method for being detected metal ion based on polymerase isothermal amplification signal amplification technique, is characterized in that
DNA enzymatic will be contained and the magnetic nano-particle surface that gold is wrapped up can be fixed to by Au-S keys from nucleic acid chains S1 of hybridization partly；Nickel
Ion（Ni2+）In the presence of, the DNA enzymatic site of nucleic acid chains S1 is by Ni2+Cutting, the excalation at 5 ' ends of nucleic acid chains S1 cause 3 '
End nucleic acid chains become single-stranded, and the single-stranded S2 of complementary seriess DNA are hybridized with S1 single stranded portions；After polymerase and substrate dNTP is added,
Under the induction progradation of S2 promote S1 to open and isothermal duplication iodine occurs, finally methylene blue MB is embedded into double
The amplification of electrochemical response signal is realized by magnetite gathering on golden carbon electrode surface in chain DNA space, the complex of formation；According to
The Ni in solution is realized in the enhancing of electrochemical signals2+The measure of concentration, comprises the following steps：
（1）Fe3O4The preparation of@Au core-shell structure copolymer magnetic nano-particles
Fe is prepared in the basic conditions initially with coprecipitation3O4Magnetic nano-particle, by 3- aminopropyl triethoxysilanes
Amination is carried out to its surface；Afterwards using reduction of sodium citrate HAuCl4×4H2O aqueous solutions, in Fe3O4Particle surface is formed
Golden shell, obtains Fe3O4@Au core-shell structure copolymer magnetic nano-particles；
（2）Double-stranded DNA flag F e3O4The preparation of@Au particles
By Fe3O4@Au particles react 12 h at 37 DEG C with nucleic acid chains S1, hatch, and Magneto separate obtains the Fe of S1 labellings3O4@Au；
By Ni2+Fe with S1 labellings3O4@Au hatch 1 h at 37 DEG C, and the DNA enzymatic site of nucleic acid chains S1 is by Ni2+Cutting, the 5 ' ends of S1
Excalation cause 3 ' end nucleic acid chains to become single-stranded, heat 5 min in 95 DEG C, nucleic acid chains S2 and S1 single stranded portions are in 37 afterwards
DEG C hybridization 3 H-shapeds into double-stranded DNA, obtain the Fe of double-stranded DNA labelling3O4@Au magnetic nano-particles；
（3）Polymerase isothermal amplification
The Fe of double-stranded DNA labelling3O4In@Au solution, at 37 DEG C, polymerase and substrate dNTP is added to react 2 h, in the induction of S2
Promote S1 to open and isothermal duplication iodine occurs under progradation, finally 3 h are reacted with methylene blue MB at 37 DEG C, make MB
It is embedded into double-stranded DNA space, the complex of formation realizes electrochemical response signal on golden carbon electrode surface by magnetite gathering
Amplify；
（4）Analysis detection
The detection that electrochemical response signal is carried out using electrochemical workstation, molten to standard according to the size of electrochemical response signal
Ni in liquid or sample solution2+Concentration is quantitative determined.
2. according to claim 1 a kind of metal ion is detected based on polymerase isothermal amplification signal amplification technique
Electrochemical method, it is characterised in that described nanoparticle is Fe3O4@Au core-shell structure copolymer magnetic nano-particles.
3. according to claim 1 a kind of metal ion is detected based on polymerase isothermal amplification signal amplification technique
Electrochemical method, it is characterised in that electrochemical signals amplifying technique is based on Fe3O4@Au core-shell structure copolymers magnetic nano-particles and polymerization
Enzyme isothermal amplification dual amplification technology.
4. according to claim 1 a kind of metal ion is detected based on polymerase isothermal amplification signal amplification technique
Electrochemical method, it is characterised in that described electrode is golden carbon electrode.
5. according to claim 1 a kind of metal ion is detected based on polymerase isothermal amplification signal amplification technique
Electrochemical method, it is characterised in that described electrochemical method is Differential Pulse Voltammetry.
6. according to claim 1 a kind of metal ion is detected based on polymerase isothermal amplification signal amplification technique
Electrochemical method, it is characterised in that described DNA sequence is as follows：
S1：5 '-ACT CAC TAT RA GGA AGA GAT GGA CGT GAG TCG ACT AGA CAC GTC CAT CTC
TGC AGT CGG GTA GTT AAA CCG ACC TTC AGA CAT AGT GAG TAG CA -3’-SH；
S2：5’-CTA CTC ACT ATG TCT-3’.
7. according to claim 1 a kind of metal ion is detected based on polymerase isothermal amplification signal amplification technique
Electrochemical method, it is characterised in that described isothermal amplification is to use electroactive material methylene blue（MB）As indicator.
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| CN113406163A (en) * | 2021-06-15 | 2021-09-17 | 国家能源集团科学技术研究院有限公司 | Magnetic induction self-assembly electrochemical biosensor for sensitively detecting trace nickel ions and application thereof |
| CN114563459A (en) * | 2022-01-27 | 2022-05-31 | 宁波大学 | Design and application research of 'binary' biological logic gate based on zinc oxide nanoparticles |
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| CN113406163A (en) * | 2021-06-15 | 2021-09-17 | 国家能源集团科学技术研究院有限公司 | Magnetic induction self-assembly electrochemical biosensor for sensitively detecting trace nickel ions and application thereof |
| CN114563459A (en) * | 2022-01-27 | 2022-05-31 | 宁波大学 | Design and application research of 'binary' biological logic gate based on zinc oxide nanoparticles |
| CN114563459B (en) * | 2022-01-27 | 2023-12-15 | 宁波大学 | Binary biological logic gate design and application research based on zinc oxide nano particles |
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