CN109959691A - A method of based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid - Google Patents
A method of based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid Download PDFInfo
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Abstract
The method based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid that this patent discloses a kind of, forms cascade structure in electrode face finish semiconductor ZnO nano material and different-grain diameter CdTe quantum, greatly enhances photo-current intensity.By the formation and dismantling of triple helical molecule, the superiority of triple helical molecule switch is that the high specific detection of nucleic acid opens new strategy, realizes the super sensitivity detection to nucleic acid by electrochemical workstation.The present invention passes through the sequence for changing adapted dna, and the photic electrochemical sensor of invention can be also used for the detection of various nucleic acid, and this method has the advantages that inexpensive, highly sensitive, response quickly, with important application prospects in bioanalysis and clinical diagnosis.
Description
Technical field
The present invention relates to a kind of detection methods of at low cost, easy to operate, high sensitivity nucleic acid, belong to the detection of nucleic acid
Technical field.
Background technique
Nucleic acid is carrier of genetic information important in organism, in terms of it stores in vital movement and propagates hereditary information
It plays a role, and determines a series of main biological phenomenas, such as the growth of biology, heredity and variation.Therefore, accurately, quickly and
The nucleic acid determination of specificity has become and is used for gene therapy in modern life science, and the core of clinical diagnosis and cancer research is asked
Topic.
So far, there are electrochemical process, Electrochemiluminescince, fluorescence radiation method and colorimetric for the method for detection of nucleic acids
Method, in the method for these detection nucleic acid, although they are each advantageous, there is also at high cost, analysis time is long, the device is complicated
The problems such as.Therefore, there is an urgent need to develop an effective platform, can not only solve the above problems, and may be implemented it is reliable and
Portable detection of nucleic acids device.
In order to improve the sensitivity of detection of nucleic acids, establishes one kind and opened based on cascade electrooptic active material and triple helical molecule
Inexpensive, highly sensitive, the quick photic electrochemica biological sensor of pass.Noble metal nano is grown using micro-fluidic paper base platform
Material increases the specific surface area of paper electrode, effectively increases the electric conductivity of electrode.Using semiconductor ZnO nano material and
Different-grain diameter CdTe quantum dot forms cascade structure, due to its unique physicochemical properties and photoelectric activity, greatly increases
Strong photo-current intensity, while also having many advantages, such as good biocompatibility, it is easily prepared, so that before it has and preferably applies
Scape.By the formation and dismantling of triple helical molecule, the superiority of triple helical molecule switch is that the high specific detection of nucleic acid is opened up
New strategy, realizes the super sensitivity detection to nucleic acid.The nucleic acid detection method established be it is a kind of with universality can
It is with important application prospects in bioanalysis and clinical diagnosis with the detection for various nucleic acid.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of designs rationally, low in cost, easy to operate, environmental-friendly
And the biosensor of high sensitivity detects nucleic acid.
A kind of method process based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid is as follows:
(1) hydrophilic region and hydrophobic wax print area of paper chip working lining and auxiliary layer are designed, it is hydrophilic in paper chip working lining
The working electrode of region preparation growth gold nanoparticle, auxiliary layer hydrophilic region prepare reference electrode and to electrode (attached drawings 1)；
(2) by the CdTe-COOH quantum dot (CdTe-COOH QDs) of ZnO nanometer rods (ZnO NRs) and two kinds of different-grain diameters
And CdTe-NH2Quantum dot (CdTe-NH2 QDs it) forms cascade light active material and is fixed on working electrode system obtained by step (1)
Standby photic electric sensing interface；
(3) DNA of hairpin structure (H-DNA) is fixed on photic electric sensing interface obtained by step (2), then uses 6- mercapto
Base hexanol (MCH) closes nonspecific activity site；
(4) alkaline phosphatase and the gold nanoparticle (ALP-Au NPs-DNA) of adapted dna modification will be marked to be fixed on step
(3) the photic electric sensing interface of gained prepares photic electrochemical sensor (attached drawing 2)；
(5) target dna (T-DNA) is added drop-wise to photic electrochemical sensor obtained by step (4), is swashed using 500W xenon source
Hair, the photoelectric current that electrochemical workstation detection generates.
A kind of method specific steps based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid
It is as follows:
(1) paper material of the present invention is chromatographic paper, and paper chip pattern is set with Adobe Illustrator CS4 software
Meter, the micro-fluidic paper chip include working lining and auxiliary layer, and border circular areas is hydrophilic region among working lining and auxiliary layer, remaining
Part is that wax prints hydrophobic region, and wherein working lining hydrophilic region diameter dimension is 6 mm, auxiliary layer hydrophilic region diameter dimension
For 8 mm；Using the method for silk-screen printing, carbon working electrode, the printing of auxiliary layer hydrophilic region are printed in working lining hydrophilic region
Ag/AgCl reference electrode and carbon are to electrode；
The step of working electrode of the present invention that growth gold nanoparticle is prepared in paper chip working lining hydrophilic region are as follows: first
80 mL distilled water are heated to 90 DEG C, the chlorauric acid solution that 0.8 mL mass fraction is 1% is then added and continues to be heated to
96 DEG C are kept for 1 minute, are eventually adding the sodium citrate that 2.8 mL mass fractions are 1%, it is molten that heating obtains gold seeds in 8 minutes
Liquid takes 20 μ L gold seeds solution to be added dropwise in working lining hydrophilic region, and standing is dried, and in triplicate, growth Jenner is prepared
The working electrode of rice corpuscles.
(2) the CdTe-COOH QDs and CdTe-NH of the present invention by ZnO NRs and two kinds of different-grain diameters2
QDs is fixed on the step of step (1) resulting working electrode in succession are as follows:
The preparation step of ZnO NRs is as follows: 0.44 g zinc acetate being dissolved in 10 mL ethanol amines to be formed under stiring
5 mL water and 15 mL ethylene glycol are added into solution and are ultrasonically treated 30 minutes, then transfer the solution into for clear solution
It is heated 4 hours in the autoclave of 100 mL polytetrafluoroethyllining linings at 180 DEG C, by its natural cooling at room temperature after reaction
To room temperature, sediment is centrifuged and is rinsed respectively 3 times with dehydrated alcohol and distilled water, is dried at room temperature for obtaining ZnO in 12 hours
Powder；
The preparation step of CdTe-COOH QDs is as follows: 89 μ L 3- mercaptopropionic acids are added to equipped with condenser and thermometer
120 mL 5mM CdCl in three-neck flask2 In aqueous solution, solution nitrogen bubbling is continued 30 minutes later, under stiring
1.0 M NaOH solutions are added dropwise and adjust pH to 11.8, then infuse 0.12 g sodium borohydride and 0.0133 g sodium tellurite in succession
Enter in solution, reaction carries out 6 hours in 100 DEG C and nitrogen atmosphere, obtains CdTe-COOH QDs solution；
CdTe-NH2The preparation step of QDs is as follows: the mercaptoethylmaine of 3.6 mmol being added in round-bottomed flask under stiring
100 mL, 12 mM CdCl2 In aqueous solution, solution high-purity nitrogen is deaerated 30 minutes, it is molten that dilute NaOH is then added
The pH of solution is adjusted to 5.7 by liquid, then injects 0.1 g sodium borohydride and 0.0133 g sodium tellurite in solution in succession,
Reaction mixture solution is heated to 100 DEG C and is flowed back 1 hour in a nitrogen atmosphere, CdTe-NH is finally obtained2QDs is molten
Liquid；
ZnO NRs is dissolved in 0.1 mM 4- aminothiophenol (PATP) ethanol solution, stirs the mixture for 4 hours, so
Sediment is centrifuged afterwards and is dissolved in ethanol solution, obtains ZnO/PATP, be added dropwise 10 ~ 200 μ L ZnO/PATP solution in
On the working electrode for growing gold nanoparticle, 20 minutes are stood at room temperature, after its drying, 20 μ L CdTe- are gradually added dropwise
COOH QDs and 20 μ L CdTe-NH2QDs is incubated for 30 minutes at room temperature, finally, using the above-mentioned work electricity of milli-Q water
Pole 3 times is dried 30 minutes at room temperature, CdTe-NH is prepared2/ CdTe-COOH/ZnO NRs cascade electrooptic active material
The photic electric sensing interface of modification.
(3) of the present invention that H-DNA is fixed on the resulting photic electric sensing interface of step (2), then sealed using MCH
The step of closing nonspecific activity site are as follows: by 20 μ L, 20 mg/mL N-(3- dimethylaminopropyl)-N'- ethyl carbon two
The mixed solution of inferior amine salt hydrochlorate and 10 mg/mL hydroxysuccinimides is added drop-wise to the resulting photic electric sensing interface of step (2)
And be incubated for 30 minutes, the H-DNA of 1 μM of 20 μ L is then added dropwise, is incubated for 16 hours, 20 μ L MCH are added and place 30 minutes envelopes
Black out sends a telegraph the nonspecific activity site of sensing interface.
(4) the step of ALP-Au NPs-DNA is fixed on photic electric sensing interface obtained by step (3) of the present invention
Are as follows: 0.2 M K is added dropwise in 2.0 mL Au NPs solution2CO3PH is adjusted to 8.2 by solution, then by 20 μ L, 1 μM of DNA
It is added thereto, is incubated for 2 hours with 40 μ L, 0.8 mg/mL alkaline phosphatase, mixture is centrifuged 20 minutes with 10000 rpm,
And washed three times with phosphoric acid washing buffer, sediment is dispersed in 200 μ L distilled water to obtain ALP-Au NPs-DNA
Dispersion liquid；It takes 20 μ L ALP-Au NPs-DNA to be added drop-wise to photic electric sensing interface, is incubated at room temperature 90 minutes, is prepared into
To photic electrochemical sensor.
(5) the step of T-DNA is added drop-wise to step (4) detection photoelectric current of the present invention are as follows: by 20 μ L T-DNA
It is added drop-wise to the working region of the resulting photic electrochemical sensor of step (4), the white light generated using 500W xenon lamp is as light source
Excitation carries out photoelectric current inspection using electrochemical workstation in the phosphate buffer solution containing 0.1 M STAY-C 50
It surveys.
Beneficial effects of the present invention
(1) present invention is using paper electrode as working electrode, and compared to traditional glass-carbon electrode and conductive glass electrode, price is low
Honest and clean, environmental protection is disposable.
(2) level link that the present invention is formed using the CdTe QDs photoelectric activity material of ZnO NRs and different-grain diameter
Structure effectively enhances photo-current intensity.
(3) present invention uses three helical dna structures, compared to traditional duplex DNA structure, has preferably spirit
Active and operability realizes the super sensitivity detection of nucleic acid by the formation and disassembly of triple helical molecule.
(4) method of detection nucleic acid of the present invention is a kind of inspection that can be used for various nucleic acid with universality
It surveys.
Detailed description of the invention
Fig. 1: the design of paper chip.
Fig. 2: the building of photic electrochemical sensor.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is further explained, but it is of the invention
Content is not limited solely to following implementation.
Embodiment
A kind of method answering in HIV-1 based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid
With, it is characterized in that the following steps are included:
(1) use the micro-fluidic paper chip of Adobe Illustrator CS4 software design working lining and auxiliary layer, working lining and
Border circular areas is hydrophilic region among auxiliary layer, and rest part is that wax prints hydrophobic region, wherein working lining hydrophilic region diameter
Having a size of 6 mm, auxiliary layer hydrophilic region diameter dimension is 8 mm；Using the method for silk-screen printing, printed in working lining hydrophilic region
Brush carbon working electrode, auxiliary layer hydrophilic region print Ag/AgCl reference electrode and carbon to electrode；
The step of working electrode of the present invention that growth gold nanoparticle is prepared in paper chip working lining hydrophilic region are as follows: first
80 mL distilled water are heated to 90 DEG C, the chlorauric acid solution that 0.8 mL mass fraction is 1% is then added and continues to be heated to
96 DEG C are kept for 1 minute, are eventually adding the sodium citrate that 2.8 mL mass fractions are 1%, it is molten that heating obtains gold seeds in 8 minutes
Liquid takes 20 μ L gold seeds solution to be added dropwise in working lining hydrophilic region, and standing is dried, and in triplicate, growth Jenner is prepared
The working electrode of rice corpuscles.
(2) the CdTe-COOH QDs and CdTe-NH of the present invention by ZnO NRs and two kinds of different-grain diameters2
QDs is fixed on the step of step (1) resulting working electrode in succession are as follows:
The preparation step of ZnO NRs is as follows: 0.44 g zinc acetate being dissolved in 10 mL ethanol amines to be formed under stiring
5 mL water and 15 mL ethylene glycol are added into solution and are ultrasonically treated 30 minutes, then transfer the solution into for clear solution
It is heated 4 hours in the autoclave of 100 mL polytetrafluoroethyllining linings at 180 DEG C, by its natural cooling at room temperature after reaction
To room temperature, sediment is centrifuged and is rinsed respectively 3 times with dehydrated alcohol and distilled water, is dried at room temperature for obtaining ZnO in 12 hours
Powder；
The preparation step of CdTe-COOH QDs is as follows: 89 μ L 3- mercaptopropionic acids are added to equipped with condenser and thermometer
120 mL 5mM CdCl in three-neck flask2 In aqueous solution, solution nitrogen bubbling is continued 30 minutes later, under stiring
1.0 M NaOH solutions are added dropwise and adjust pH to 11.8, then infuse 0.12 g sodium borohydride and 0.0133 g sodium tellurite in succession
Enter in solution, reaction carries out 6 hours in 100 DEG C and nitrogen atmosphere, obtains CdTe-COOH QDs solution；
CdTe-NH2The preparation step of QDs is as follows: the mercaptoethylmaine of 3.6 mmol being added in round-bottomed flask under stiring
100 mL, 12 mM CdCl2 In aqueous solution, solution high-purity nitrogen is deaerated 30 minutes, it is molten that dilute NaOH is then added
The pH of solution is adjusted to 5.7 by liquid, then injects 0.1 g sodium borohydride and 0.0133 g sodium tellurite in solution in succession,
Reaction mixture solution is heated to 100 DEG C and is flowed back 1 hour in a nitrogen atmosphere, CdTe-NH is finally obtained2QDs is molten
Liquid；
ZnO NRs is dissolved in 0.1 mM 4- aminothiophenol (PATP) ethanol solution, stirs the mixture for 4 hours, so
Sediment is centrifuged afterwards and is dissolved in ethanol solution, obtains ZnO/PATP, be added dropwise 10 ~ 200 μ L ZnO/PATP solution in
On the working electrode for growing gold nanoparticle, 20 minutes are stood at room temperature, after its drying, 20 μ L CdTe- are gradually added dropwise
COOH QDs and 20 μ L CdTe-NH2QDs is incubated for 30 minutes at room temperature, finally, using the above-mentioned work electricity of milli-Q water
Pole 3 times is dried 30 minutes at room temperature, CdTe-NH is prepared2/ CdTe-COOH/ZnO NRs cascade electrooptic active material
The photic electric sensing interface of modification.
(3) of the present invention that H-DNA is fixed on the resulting photic electric sensing interface of step (2), then sealed using MCH
The step of closing nonspecific activity site are as follows: by 20 μ L, 20 mg/mL N-(3- dimethylaminopropyl)-N'- ethyl carbon two
The mixed solution of inferior amine salt hydrochlorate and 10 mg/mL hydroxysuccinimides is added drop-wise to the resulting photic electric sensing interface of step (2)
And be incubated for 30 minutes, the H-DNA of 1 μM of 20 μ L is then added dropwise, is incubated for 16 hours, 20 μ L MCH are added and place 30 minutes envelopes
Black out sends a telegraph the nonspecific activity site of sensing interface.
(4) the step of ALP-Au NPs-DNA is fixed on photic electric sensing interface obtained by step (3) of the present invention
Are as follows: 0.2 M K is added dropwise in 2.0 mL Au NPs solution2CO3PH is adjusted to 8.2 by solution, then by 20 μ L, 1 μM of DNA
It is added thereto, is incubated for 2 hours with 40 μ L, 0.8 mg/mL alkaline phosphatase, mixture is centrifuged 20 minutes with 10000 rpm,
And washed three times with phosphoric acid washing buffer, sediment is dispersed in 200 μ L distilled water to obtain ALP-Au NPs-DNA
Dispersion liquid；It takes 20 μ L ALP-Au NPs-DNA to be added drop-wise to photic electric sensing interface, is incubated at room temperature 90 minutes, is prepared into
To photic electrochemical sensor.
(5) HIV-1 is detected: 20 μ L HIV-1-DNA are added drop-wise to the resulting photic electrochemical sensor of step (4)
Working region, the white light generated using 500W xenon lamp is as light source activation, in the phosphoric acid containing 0.1 M STAY-C 50
Photoelectric current detection is carried out using electrochemical workstation in salt buffer solution.
Sequence table
<110>University Of Ji'nan
<120>a kind of method based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid
<130> 2019
<141> 2019-04-11
<160> 3
<170> SIPOSequenceListing 1.0
<210> 4
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
gcagtgagag aggagactgc 20
<210> 4
<211> 35
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
ctctctcatg tggaaaatct ctagcagtct ctctc 35
<210> 4
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
actgctagag attttccaca t 21
Claims (6)
1. a kind of method based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid, feature includes following step
It is rapid:
(1) hydrophilic region and hydrophobic wax print area of paper chip working lining and auxiliary layer are designed, it is hydrophilic in paper chip working lining
The working electrode of region preparation growth gold nanoparticle, auxiliary layer hydrophilic region prepare reference electrode and to electrodes；
(2) by the CdTe-COOH quantum dot (CdTe-COOH QDs) of ZnO nanometer rods (ZnO NRs) and two kinds of different-grain diameters
And CdTe-NH2Quantum dot (CdTe-NH2 QDs it) forms cascade electrooptic active material and is fixed on working electrode obtained by step (1)
Prepare photic electric sensing interface；
(3) DNA of hairpin structure (H-DNA) is fixed on photic electric sensing interface obtained by step (2), then uses 6- mercapto
Base hexanol (MCH) closes nonspecific activity site；
(4) alkaline phosphatase and the gold nanoparticle (ALP-Au NPs-DNA) of adapted dna modification will be marked to be fixed on step
(3) the photic electric sensing interface of gained prepares photic electrochemical sensor；
(5) target dna (T-DNA) is added drop-wise to photic electrochemical sensor obtained by step (4), is swashed using 500W xenon source
Hair, the photoelectric current that electrochemical workstation detection generates.
2. a kind of side based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid according to claims 1
Method, which is characterized in that paper material as described in step (1) is chromatographic paper, paper chip pattern Adobe Illustrator
CS4 software design, the micro-fluidic paper chip include working lining and auxiliary layer, and border circular areas is parent among working lining and auxiliary layer
Water area, rest part are that wax prints hydrophobic region, and wherein working lining hydrophilic region diameter dimension is 6 mm, auxiliary layer hydrophilic area
Domain diameter dimension is 8 mm；Using the method for silk-screen printing, carbon working electrode is printed in working lining hydrophilic region, auxiliary layer is hydrophilic
Ag/AgCl reference electrode and carbon are printed to electrode in region；
The step of working electrode of the present invention that growth gold nanoparticle is prepared in paper chip working lining hydrophilic region are as follows: first
80 mL distilled water are heated to 90 DEG C, the chlorauric acid solution that 0.8 mL mass fraction is 1% is then added and continues to be heated to
96 DEG C are kept for 1 minute, are eventually adding the sodium citrate that 2.8 mL mass fractions are 1%, it is molten that heating obtains gold seeds in 8 minutes
Liquid takes 20 μ L gold seeds solution to be added dropwise in working lining hydrophilic region, and standing is dried, and in triplicate, growth Jenner is prepared
The working electrode of rice corpuscles.
3. a kind of side based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid according to claims 1
Method, which is characterized in that by the CdTe-COOH QDs and CdTe-NH of ZnO NRs and two kinds of different-grain diameters2QDs is successive
The step of being fixed on step (1) resulting working electrode are as follows:
The preparation step of ZnO NRs is as follows: 0.44 g zinc acetate being dissolved in 10 mL ethanol amines to be formed under stiring
5 mL water and 15 mL ethylene glycol are added into solution and are ultrasonically treated 30 minutes, then transfer the solution into for clear solution
It is heated 4 hours in the autoclave of 100 mL polytetrafluoroethyllining linings at 180 DEG C, by its natural cooling at room temperature after reaction
To room temperature, sediment is centrifuged and is rinsed respectively 3 times with dehydrated alcohol and distilled water, is dried at room temperature for obtaining ZnO in 12 hours
Powder；
The preparation step of CdTe-COOH QDs is as follows: 89 μ L 3- mercaptopropionic acids are added to equipped with condenser and thermometer
120 mL 5mM CdCl in three-neck flask2 In aqueous solution, solution nitrogen bubbling is continued 30 minutes later, under stiring
1.0 M NaOH solution are added dropwise and adjust pH to 11.8, it is then that 0.12 g sodium borohydride and 0.0133 g sodium tellurite is successive
It injects in solution, reaction carries out 6 hours in 100 DEG C and nitrogen atmosphere, obtains CdTe-COOH QDs solution；
CdTe-NH2The preparation step of QDs is as follows: the mercaptoethylmaine of 3.6 mmol being added in round-bottomed flask under stiring
100 mL, 12 mM CdCl2 In aqueous solution, solution high-purity nitrogen is deaerated 30 minutes, it is molten that dilute NaOH is then added
The pH of solution is adjusted to 5.7 by liquid, then injects 0.1 g sodium borohydride and 0.0133 g sodium tellurite in solution in succession,
Reaction mixture solution is heated to 100 DEG C and is flowed back 1 hour in a nitrogen atmosphere, CdTe-NH is finally obtained2QDs is molten
Liquid；
ZnO NRs is dissolved in 0.1 mM 4- aminothiophenol (PATP) ethanol solution, stirs the mixture for 4 hours, so
Sediment is centrifuged afterwards and is dissolved in ethanol solution, obtains ZnO/PATP, be added dropwise 10 ~ 200 μ L ZnO/PATP solution in
On the working electrode for growing gold nanoparticle, 20 minutes are stood at room temperature, after its drying, 20 μ L CdTe- are gradually added dropwise
COOH QDs and 20 μ L CdTe-NH2QDs is incubated for 30 minutes at room temperature, finally, using the above-mentioned work electricity of milli-Q water
Pole 3 times is dried 30 minutes at room temperature, CdTe-NH is prepared2/ CdTe-COOH/ZnO NRs cascade electrooptic active material
The photic electric sensing interface of modification.
4. a kind of side based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid according to claims 1
Method, which is characterized in that H-DNA is fixed on the resulting photic electric sensing interface of step (2), non-spy is then closed using MCH
The step of specific activities site are as follows: by 20 μ L, 20 mg/mL N-(3- dimethylaminopropyl)-N'- ethyl carbodiimide salt
The mixed solution of hydrochlorate and 10 mg/mL hydroxysuccinimides is added drop-wise to the resulting photic electric sensing interface of step (2) and is incubated for
30 minutes, the H-DNA of 1 μM of 20 μ L is then added dropwise, is incubated for 16 hours, it is photic that 20 μ L MCH placement closing in 30 minutes is added
The nonspecific activity site of electric sensing interface.
5. a kind of side based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid according to claims 1
Method, which is characterized in that the step of ALP-Au NPs-DNA is fixed on photic electric sensing interface obtained by step (3) are as follows: 2.0
0.2 M K is added dropwise in mL Au NPs solution2CO3PH is adjusted to 8.2 by solution, then by 20 μ L 1 μM of DNA and 40 μ L
0.8 mg/mL alkaline phosphatase is added thereto, and is incubated for 2 hours, by mixture with 10000 rpm centrifugation 20 minutes, and uses phosphoric acid
Washing buffer is washed three times, and sediment is dispersed in 200 μ L distilled water to obtain ALP-Au NPs-DNA dispersion liquid；
It takes 20 μ L ALP-Au NPs-DNA to be added drop-wise to photic electric sensing interface, is incubated at room temperature 90 minutes, photic electricity is prepared
Chemical sensor.
6. a kind of side based on cascade electrooptic active material and triple helical molecule switch detection nucleic acid according to claims 1
Method, which is characterized in that the step of T-DNA is added drop-wise to step (4) detection photoelectric current are as follows: 20 μ L T-DNA are added drop-wise to step
Suddenly the working region of (4) resulting photic electrochemical sensor, the white light generated using 500W xenon lamp are being contained as light source activation
Have in the phosphate buffer solution of 0.1 M STAY-C 50 and carries out photoelectric current detection using electrochemical workstation.
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| CN110274948A (en) * | 2019-07-11 | 2019-09-24 | 青岛科技大学 | One kind is based on the bis- amplification ECL biosensors of triple helical molecule switch oversoul sensitive detection LPS and its application |
| CN110320258A (en) * | 2019-07-04 | 2019-10-11 | 青岛科技大学 | A method of nucleic acid molecules are detected based on cyclic amplification and cation exchange |
| CN110376260A (en) * | 2019-07-18 | 2019-10-25 | 济南大学 | Detect the preparation method of the optical electro-chemistry aptamer sensor of prostate-specific antigen |
| CN110530950A (en) * | 2019-09-26 | 2019-12-03 | 济南大学 | The building of paper base sensor based on exciton Plasma Interaction efficient detection PSA |
| CN111304297A (en) * | 2020-03-23 | 2020-06-19 | 江苏师范大学 | Method for analyzing circulating tumor DNA on single molecule level |
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| CN110376260B (en) * | 2019-07-18 | 2021-11-02 | 济南大学 | Preparation method of photoelectrochemical aptamer sensor for detecting prostate specific antigen |
| CN110530950A (en) * | 2019-09-26 | 2019-12-03 | 济南大学 | The building of paper base sensor based on exciton Plasma Interaction efficient detection PSA |
| CN111304297A (en) * | 2020-03-23 | 2020-06-19 | 江苏师范大学 | Method for analyzing circulating tumor DNA on single molecule level |
| CN112098484A (en) * | 2020-09-11 | 2020-12-18 | 常州大学 | Sensor for detecting acetamiprid based on electrochemical luminescence method, preparation method and application |
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