CN103913500B - A kind of based on TiO 2fe 3o 4the dendrimers electrochemica biological sensor, its preparation method and the purposes that form with DNA - Google Patents
A kind of based on TiO 2fe 3o 4the dendrimers electrochemica biological sensor, its preparation method and the purposes that form with DNA Download PDFInfo
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- CN103913500B CN103913500B CN201410166673.4A CN201410166673A CN103913500B CN 103913500 B CN103913500 B CN 103913500B CN 201410166673 A CN201410166673 A CN 201410166673A CN 103913500 B CN103913500 B CN 103913500B
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- 239000000412 dendrimer Substances 0.000 title claims abstract description 39
- 229920000736 dendritic polymer Polymers 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 108020004414 DNA Proteins 0.000 claims abstract description 74
- 229910010413 TiO 2 Inorganic materials 0.000 claims abstract description 69
- 230000000694 effects Effects 0.000 claims abstract description 23
- 102000053602 DNA Human genes 0.000 claims abstract description 15
- 238000001514 detection method Methods 0.000 claims abstract description 11
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 9
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 31
- 239000010931 gold Substances 0.000 claims description 31
- 229910052737 gold Inorganic materials 0.000 claims description 31
- 239000002131 composite material Substances 0.000 claims description 26
- 239000007853 buffer solution Substances 0.000 claims description 25
- 239000000523 sample Substances 0.000 claims description 17
- 239000000126 substance Substances 0.000 claims description 12
- 239000002105 nanoparticle Substances 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 10
- 238000004506 ultrasonic cleaning Methods 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- 230000026731 phosphorylation Effects 0.000 claims description 9
- 238000006366 phosphorylation reaction Methods 0.000 claims description 9
- 238000005498 polishing Methods 0.000 claims description 9
- 238000006555 catalytic reaction Methods 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- 238000001338 self-assembly Methods 0.000 claims description 8
- 239000011248 coating agent Substances 0.000 claims description 6
- 238000000576 coating method Methods 0.000 claims description 6
- 238000001027 hydrothermal synthesis Methods 0.000 claims description 6
- 239000004411 aluminium Substances 0.000 claims description 5
- 229910052782 aluminium Inorganic materials 0.000 claims description 5
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- 230000033444 hydroxylation Effects 0.000 claims description 5
- 238000005805 hydroxylation reaction Methods 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 5
- 239000012498 ultrapure water Substances 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 239000002114 nanocomposite Substances 0.000 claims description 4
- 230000000295 complement effect Effects 0.000 abstract description 3
- 230000009870 specific binding Effects 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 238000012546 transfer Methods 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 2
- 238000002484 cyclic voltammetry Methods 0.000 description 8
- 238000010586 diagram Methods 0.000 description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000004381 surface treatment Methods 0.000 description 3
- 108091023037 Aptamer Proteins 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
- 238000007306 functionalization reaction Methods 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000002122 magnetic nanoparticle Substances 0.000 description 1
- 239000002082 metal nanoparticle Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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Abstract
The present invention relates to a kind of based on TiO
2fe
3o
4the dendrimers electrochemica biological sensor formed with DNA, its preparation method and purposes, use water heat transfer TiO
2fe
3o
4nano-complex, utilizes one end phosphorylated cdna and TiO
2between specific binding effect, utilize the complementary pairing effect of base between DNA sequence dna, prepare based on TiO
2fe
3o
4with the dendrimers electrochemica biological sensor of DNA, this sensor achieves the detection to target dna molecule sensitivity, specificity, stability.
Description
Technical field
The invention belongs to biosensor technology field, be specifically related to use TiO
2fe
3o
4the electrochemica biological sensor that the dendrimers formed with DNA is assembled and be applied to the detection of target dna molecule.
Background technology
21 century is century of life science, and be an extremely important aspect in life science to the research of DNA, the research of DNA biosensor becomes focus.Research much related to this is had to be devoted to amplify with metal nanoparticle the measured signal of DNA biosensor.Wherein what is interesting is and find Fe
3o
4magnetic nano-particle, it not only can be widely used in bio-imaging technology, and there is the performance of similar peroxidase activity catalysis, and can by other nano material functionalization coated catalysts or enzyme, thus make the function nano compound prepared can be used to research better in conjunction with its nucleocapsid function.
Summary of the invention
The object of the present invention is to provide a kind of based on TiO
2fe
3o
4the dendrimers electrochemica biological sensor formed with DNA, its preparation method and purposes, utilize the base complementrity effect of DNA in target dna molecule and dendrimers, at the surface-assembled electrochemica biological sensor of gold electrode, be applied to the detection of target dna molecule.Concrete technical scheme is as follows:
A kind of based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, comprises the steps:
(1) DNA sequence dna: sulfhydrylation: S1, target dna: S2, one end hydroxylation: S3, be dissolved in buffer solution respectively, and save backup;
(2) buffer solution containing probe S1 is dripped be coated onto on gold electrode, cultivate, the gold electrode modified by Au-S covalent bond effect self assembly S1;
(3) with step (2) simultaneously, in the buffer solution containing probe S3, add T4PNK, ATP and cultivate this mixed solution, obtaining the S3 of phosphorylation;
(4) TiO prepared is added
2fe
3o
4compound substance, continues to cultivate, obtains DNA-TiO
2fe
3o
4biological composite;
(5) gold electrode that S1 modifies is put into containing S2 and DNA-TiO
2fe
3o
4cultivate in the buffer solution of biological composite, utilize the base pair complementarity effect between DNA, based on TiO
2fe
3o
4the dendrimers formed with DNA is assembled electrochemica biological sensor and is successfully prepared.
Further, TiO described in step (4)
2fe
3o
4the preparation of nano composite material comprises the steps:
(4-1) utilize hydrothermal synthesis method, synthesize the Fe of size uniform
3o
4nano particle;
(4-2) Fe will be successfully prepared
3o
4nano particle utilizes hydrothermal synthesis method at its Surface coating one deck TiO
2coating;
(4-3) compound substance syncaryon Fe
3o
4living things catalysis performance and shell TiO
2biologic specificity.
Further, in step (1), DNA sequence dna is dissolved in respectively in 0.05MTris-HCl (pH7.4) buffer solution, and saves backup at 4 DEG C.
Further, gold electrode described in step (2) carried out polishing and cleaning before step (2).
Further, described polishing and cleaning comprise the steps: gold electrode first to carry out polishing with the aluminium powder of 0.3 and 0.5 μm successively; Put into HNO successively
3: H
2in O (v/v)=1:1 solution, ethanolic solution, ultrapure water, carry out Ultrasonic Cleaning, the time of ultrasonic cleaning is respectively 3 ~ 5min.
Further, in step (2), cultivate 6-10h, the gold electrode modified by Au-S covalent bond effect self assembly S1; And/or step (3) adds T4PNK, ATP and cultivates this mixed solution 3-5h in the buffer solution containing probe S3, obtains the S3 of phosphorylation; And/or step (4) adds the TiO prepared
2fe
3o
4compound substance, continues to cultivate 1-3h, obtains DNA-TiO
2fe
3o
4biological composite.
Further, in step (2), cultivate h, the gold electrode modified by Au-S covalent bond effect self assembly S1; Step (3) adds T4PNK, ATP and cultivates this mixed solution 5h in the buffer solution containing probe S3, obtains the S3 of phosphorylation; Step (4) adds the TiO prepared
2fe
3o
4compound substance, continues to cultivate 3h, obtains DNA-TiO
2fe
3o
4biological composite.
A kind of based on TiO
2fe
3o
4the dendrimers electrochemica biological sensor formed with DNA, adopts above-mentioned preparation method to prepare.
A kind of above-mentioned based on TiO
2fe
3o
4the purposes of the dendrimers electrochemica biological sensor formed with DNA, for the detection to target dna molecule.
Further, the concentration of S2 is different, the DNA-TiO of assembling
2fe
3o
4the amount of biological composite is different, realizes variable concentrations target dna S2 and quantitatively detects.
Compared with currently available technology, the invention provides TiO
2fe
3o
4the electrochemica biological sensor that the dendrimers formed with DNA is assembled and be applied to the detection of target dna molecule, the present invention uses water heat transfer TiO
2fe
3o
4nano-complex, utilizes one end phosphorylated cdna and TiO
2between specific binding effect, utilize the complementary pairing effect of base between DNA sequence dna, prepare based on TiO
2fe
3o
4with the dendrimers electrochemica biological sensor of DNA, this sensor achieves the detection to target dna molecule sensitivity, specificity, stability.
Specifically, the preparation method of this biology sensor, the nano-complex synthesis used is simple, consume energy low, cost is low, good biocompatibility, and the biology sensor of preparation detects the signal carrying out amplifying Electrochemical Detection with the catalytic action of nano composite material, by target dna molecule and DNA-TiO
2fe
3o
4base complementrity effect between DNA in biological composite, the sensor of dendrimer structure can be prepared, can the nano-complex more with living things catalysis performance be assembled on sensor, therefore the detection to target dna molecule can be realized, it is satisfactory that result shows the testing result of this biology sensor to target dna molecule, the range of linearity detected is wider, about from 10
-10m is to 10
-6have sensitiveer detection within the scope of M, and have highly sensitive, detectability is low, selectivity good, the feature of good stability.In addition, this electrochemica biological sensor is the base pair complementarity effect utilized between DNA, by TiO
2fe
3o
4the dendrimers formed by target dna molecule connection function with DNA biological composite, and this dendrimers is assembled on electrode formed.Electrochemica biological sensor is successfully prepared.Given this, we can utilize other method of attachment (the such as base mispairing of cytimidine-silver ion-cytimidine or the specific binding etc. of aptamers and aptamer) by TiO
2fe
3o
4form dendrimers with DNA biological composite and be assembled into electrochemica biological sensor, realizing the detection to specific ion or protein.
Accompanying drawing explanation
Fig. 1 (A) is the Fe of preparation
3o
4nano particle and TiO
2fe
3o
4the scanning electron microscope (SEM) photograph of composite nanoparticle.
Fig. 1 (B) is the Fe of preparation
3o
4nano particle and TiO
2fe
3o
4composite nanoparticle infrared spectrogram.
Fig. 2 is the impedance diagram of each step electrode modification process electrode.
In figure:
A is the impedance diagram of naked gold electrode.
B is the impedance diagram of the gold electrode that probe S1 modifies.
C is probe S1 and S3-TiO
2fe
3o
4the impedance diagram of the gold electrode modified after effect.
D is the impedance diagram of the gold electrode that probe S1 and S2 and phosphorylation S3 hybridization are modified.
E is for being probe S1 and S2 and S3-TiO
2fe
3o
4the impedance diagram of the gold electrode modified after effect.
Fig. 3 (A) is based on TiO
2fe
3o
4the dendrimers formed with DNA assembles the cyclic voltammogram of electrochemica biological sensor to the catalysis of variable concentrations hydrogen peroxide.
Fig. 3 (B) for this reason sensor to the typical curve of variable concentrations hydrogen peroxide catalysis.
Fig. 4 is the cyclic voltammogram of different modifying electrode pair hydrogen peroxide catalysis.
In Fig. 4 (A):
A is the cyclic voltammogram of ordinary construction sensor.
B is based on TiO
2fe
3o
4the dendrimers formed with DNA assembles the cyclic voltammogram of electrochemica biological sensor.
In Fig. 4 (B):
A is and S3-TiO
2fe
3o
4the DNA of the random pairing cyclic voltammogram of sensor for assembling.
B is and S3-TiO
2fe
3o
4there is the cyclic voltammogram of sensor of the DNA of two base mispairings for assembling.
C is and S3-TiO
2fe
3o
4the cyclic voltammogram of sensor of the DNA having complete complementary to match for assembling.
Fig. 5 (A) is based on TiO
2fe
3o
4the dendrimers formed with DNA assembles the cyclic voltammogram that electrochemica biological sensor detects variable concentrations target dna molecule.
Fig. 5 (B) is based on TiO
2fe
3o
4the dendrimers formed with DNA assembles the chronoamperogram that electrochemica biological sensor detects variable concentrations target dna molecule.
Fig. 5 (C) is based on TiO
2fe
3o
4the dendrimers formed with DNA assembles the typical curve of electrochemica biological sensor to the change that variable concentrations target dna molecule detects.
Fig. 6 is based on TiO
2fe
3o
4the electrochemica biological sensor that the dendrimers formed with DNA is assembled prepare schematic diagram.
Embodiment
Describe the present invention with reference to the accompanying drawings below, it is a kind of preferred embodiment in numerous embodiments of the present invention.
Embodiment one:
TiO
2fe
3o
4the preparation of nano composite material: utilize hydrothermal synthesis method, first synthesizes the Fe of size uniform
3o
4nano particle, then the Fe that will be successfully prepared
3o
4nano particle utilizes hydrothermal synthesis method at its Surface coating one deck TiO
2coating, this compound substance will effective syncaryon (Fe
3o
4) living things catalysis performance and shell (TiO
2) biologic specificity performance.Therefore the detection compound substance of preparation being used for biology sensor has good using value.
Based on TiO
2fe
3o
4the step of the dendrimers electrochemica biological sensor preparations and applicatio formed with DNA is as follows:
A, the DNA sequence dna (sulfhydrylation: S1, target dna: S2, one end hydroxylation: S3) bought to be dissolved in Tris-HCl (pH7.4) buffer solution respectively, and to save backup down at low temperatures.
B, gold electrode is first carried out polishing with the aluminium powder of 0.3 and 0.5mm successively, then put into HNO successively
3: H
2in O (v/v)=1:1 solution, ethanolic solution, ultrapure water, carry out Ultrasonic Cleaning, the time of ultrasonic cleaning is respectively 3 ~ 5min.
C, the buffer solution containing probe S1 dripped be coated onto on the clean gold electrode of surface treatment, cultivate 6-10h, the gold electrode modified by Au-S covalent bond effect self assembly S1.Meanwhile, in the buffer solution containing probe S3, add T4PNK, ATP and cultivate this mixed solution 3-5h, obtaining the S3 of phosphorylation; Add the TiO prepared again
2fe
3o
4compound substance, continues to cultivate 1-3h, obtains DNA-TiO
2fe
3o
4biological composite.
D, by S1 modify gold electrode put into containing S2 and DNA-TiO
2fe
3o
4cultivate 3-5h in the buffer solution of biological composite, utilize the base pair complementarity effect between DNA, based on TiO
2fe
3o
4the dendrimers formed with DNA is assembled electrochemica biological sensor and is successfully prepared.
E, due to the concentration of S2 different, the DNA-TiO of assembling
2fe
3o
4the amount of biological composite can be different, along with the increase of S2 concentration, and DNA-TiO
2fe
3o
4the amount of biological composite also can increase thereupon, and therefore this sensor quantitatively can detect variable concentrations target dna S2.
Embodiment two:
Based on TiO
2fe
3o
4the step of the dendrimers electrochemica biological sensor preparations and applicatio formed with DNA is as follows:
A, the DNA sequence dna (sulfhydrylation: S1, target dna: S2, one end hydroxylation: S3) bought to be dissolved in 0.05MTris-HCl (pH7.4) buffer solution respectively, and to save backup at 4 DEG C.
B, gold electrode is first carried out polishing with the aluminium powder of 0.3 and 0.5mm successively, then put into HNO successively
3: H
2in O (v/v)=1:1 solution, ethanolic solution, ultrapure water, carry out Ultrasonic Cleaning, the time of ultrasonic cleaning is respectively 3 ~ 5min.
C, the buffer solution containing probe S1 dripped be coated onto on the clean gold electrode of surface treatment, cultivate 6-10h, the gold electrode modified by Au-S covalent bond effect self assembly S1.Meanwhile, in the buffer solution containing probe S3, add T4PNK, ATP and cultivate this mixed solution 3-5h, obtaining the S3 of phosphorylation; Add the TiO prepared again
2fe
3o
4compound substance, continues to cultivate 1-3h, obtains DNA-TiO
2fe
3o
4biological composite.
D, by S1 modify gold electrode put into containing S2 and DNA-TiO
2fe
3o
4cultivate 3-5h in the buffer solution of biological composite, utilize the base pair complementarity effect between DNA, based on TiO
2fe
3o
4the dendrimers formed with DNA is assembled electrochemica biological sensor and is successfully prepared.
E, due to the concentration of S2 different, the DNA-TiO of assembling
2fe
3o
4the amount of biological composite can be different, along with the increase of S2 concentration, and DNA-TiO
2fe
3o
4the amount of biological composite also can increase thereupon, and therefore this sensor quantitatively can detect variable concentrations target dna S2.
Based on TiO
2fe
3o
4the step of the dendrimers electrochemica biological sensor preparations and applicatio formed with DNA is as follows:
A, the DNA sequence dna (sulfhydrylation: S1, target dna: S2, one end hydroxylation: S3) bought to be dissolved in 0.05MTris-HCl (pH7.4) buffer solution respectively, and to save backup at 4 DEG C.
B, gold electrode is first carried out polishing with the aluminium powder of 0.3 and 0.5mm successively, then put into HNO successively
3: H
2in O (v/v)=1:1 solution, ethanolic solution, ultrapure water, carry out Ultrasonic Cleaning, the time of ultrasonic cleaning is respectively 3 ~ 5min.
C, the buffer solution containing probe S1 dripped be coated onto on the clean gold electrode of surface treatment, cultivate 10h, the gold electrode modified by Au-S covalent bond effect self assembly S1.Meanwhile, in the buffer solution containing probe S3, add T4PNK, ATP and cultivate this mixed solution 5h, obtaining the S3 of phosphorylation; Add the TiO prepared again
2fe
3o
4compound substance, continues to cultivate 3h, obtains DNA-TiO
2fe
3o
4biological composite.
D, by S1 modify gold electrode put into containing S2 and DNA-TiO
2fe
3o
4cultivate 5h in the buffer solution of biological composite, utilize the base pair complementarity effect between DNA, based on TiO
2fe
3o
4the dendrimers formed with DNA is assembled electrochemica biological sensor and is successfully prepared.
E, due to the concentration of S2 different, the DNA-TiO of assembling
2fe
3o
4the amount of biological composite can be different, along with the increase of S2 concentration, and DNA-TiO
2fe
3o
4the amount of biological composite also can increase thereupon, and therefore this sensor quantitatively can detect variable concentrations target dna S2.
Above by reference to the accompanying drawings to invention has been exemplary description; obvious specific implementation of the present invention is not subject to the restrictions described above; as long as have employed the various improvement that method of the present invention is conceived and technical scheme is carried out; or directly apply to other occasion, all within protection scope of the present invention without improving.
Claims (9)
1. one kind based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, comprise the steps:
(1) DNA sequence dna: sulfhydrylation: S1, target dna: S2, one end hydroxylation: S3, be dissolved in buffer solution respectively, and save backup;
(2) buffer solution containing probe S1 is dripped be coated onto on gold electrode, cultivate, the gold electrode modified by Au-S covalent bond effect self assembly S1;
(3) with step (2) simultaneously, in the buffer solution containing probe S3, add T4PNK, ATP and cultivate this mixed solution, obtaining the S3 of phosphorylation;
(4) TiO prepared is added
2fe
3o
4compound substance, continues to cultivate, obtains DNA-TiO
2fe
3o
4biological composite;
(5) gold electrode that S1 modifies is put into containing S2 and DNA-TiO
2fe
3o
4cultivate in the buffer solution of biological composite, utilize the base pair complementarity effect between DNA, based on TiO
2fe
3o
4the dendrimers formed with DNA is assembled electrochemica biological sensor and is successfully prepared.
2. as claimed in claim 1 based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, TiO described in step (4)
2fe
3o
4the preparation of nano composite material comprises the steps:
(4-1) utilize hydrothermal synthesis method, synthesize the Fe of size uniform
3o
4nano particle;
(4-2) Fe will be successfully prepared
3o
4nano particle utilizes hydrothermal synthesis method at its Surface coating one deck TiO
2coating;
(4-3) compound substance syncaryon Fe
3o
4living things catalysis performance and shell TiO
2biologic specificity.
3. as claimed in claim 1 or 2 based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, in step (1), DNA sequence dna being dissolved in respectively pH is in the 0.05MTris-HCl buffer solution of 7.4, and saves backup at 4 DEG C.
4. according to any one of claim 1-3 based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, gold electrode described in step (2) carried out polishing and cleaning before step (2).
5. as claimed in claim 4 based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, described polishing and cleaning comprise the steps: gold electrode first to carry out polishing with the aluminium powder of 0.3 and 0.5mm successively; Put into HNO successively
3: H
2in O (v/v)=1:1 solution, ethanolic solution, ultrapure water, carry out Ultrasonic Cleaning, the time of ultrasonic cleaning is respectively 3 ~ 5min.
6. as claimed in claim 1 or 2 based on TiO
2fe
3o
4the preparation method of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, cultivates 6-10h in step (2), the gold electrode modified by Au-S covalent bond effect self assembly S1; And/or step (3) adds T4PNK, ATP and cultivates this mixed solution 3-5h in the buffer solution containing probe S3, obtains the S3 of phosphorylation; And/or step (4) adds the TiO prepared
2fe
3o
4compound substance, continues to cultivate 1-3h, obtains DNA-TiO
2fe
3o
4biological composite.
7. one kind based on TiO
2fe
3o
4the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, adopts the preparation method described in claim 1 or 2 to prepare.
8. one kind as claimed in claim 7 based on TiO
2fe
3o
4the purposes of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, for the detection to target dna molecule.
9. as claimed in claim 8 based on TiO
2fe
3o
4the purposes of the dendrimers electrochemica biological sensor formed with DNA, is characterized in that, the concentration of S2 is different, the DNA-TiO of assembling
2fe
3o
4the amount of biological composite is different, realizes variable concentrations target dna S2 and quantitatively detects.
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| CN106501343B (en) * | 2016-10-19 | 2019-04-16 | 云南民族大学 | A method of heavy metal ion is detected based on polymerase isothermal amplification technique |
| CN112697858B (en) * | 2020-12-08 | 2022-02-18 | 云南民族大学 | Electrochemical analysis method for determining activity of T4 polynucleotide kinase based on magnetic nano material |
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