CN104330454B - A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application - Google Patents

A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application Download PDF

Info

Publication number
CN104330454B
CN104330454B CN201410601204.0A CN201410601204A CN104330454B CN 104330454 B CN104330454 B CN 104330454B CN 201410601204 A CN201410601204 A CN 201410601204A CN 104330454 B CN104330454 B CN 104330454B
Authority
CN
China
Prior art keywords
srtio
coo
composite material
nano composite
preparation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN201410601204.0A
Other languages
Chinese (zh)
Other versions
CN104330454A (en
Inventor
庞雪辉
魏琴
吴丹
范大伟
王友英
任伟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Jinan
Original Assignee
University of Jinan
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Jinan filed Critical University of Jinan
Priority to CN201410601204.0A priority Critical patent/CN104330454B/en
Publication of CN104330454A publication Critical patent/CN104330454A/en
Application granted granted Critical
Publication of CN104330454B publication Critical patent/CN104330454B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Abstract

The present invention relates to a kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application thereof, belong to bio-sensing detection technique field.Based on SrTiO 3/ CoO-WO 3compound, as beacon material, can realize the qualitative and quantitative analysis to solid tissue target dna and house-keeping gene thereof, has the advantage that equipment is simple, cost is low, be easy to microminiaturization.

Description

A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application
Technical field
The present invention relates to a kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application thereof, the detection of the biological tissue DNA that this sensor is used for, belongs to bio-sensing detection technique field.
Background technology
It is the topic that people are very popular in recent years that Gene science and DNA detect, along with the further investigation to Gene science, completing of the Human Genome Project, the mankind have cracked the secret code of life, human social development is to today, Gene science gradually grasp by people, and utilize bring benefit to the mankind self.Gene science industry is exactly the high-tech industry of the frontier nature of arising at the historic moment under this overall background.
The U.S. is the country that the whole world utilizes genetic test predictive disease the earliest, and within 2005, have nearly 5,000,000 person-times and receive detection, by precognition and medical intervention, make the incidence of disease of colorectal cancer have dropped 90%, the incidence of disease of breast cancer have dropped 70%.China relatively lags behind in genetic test, and just having an appointment 100,000 person-times by the end of the year 2006 receives detection.And China nearly 5 years cancer patients are up to 7,500,000 people, increase more than 29% less than 20 years cancer morbidities, about have 1,600,000 people to die from cancer every year.Show according to statistics, the economic loss that China causes because of disease is up to 14,000 hundred million yuan.China's gene field famous scientist's hair makes people rich and points out, genetic test is people for understanding the personal feature of gene and the detection of gene expression characteristics carried out, and the series of physiological and pathological of this mankind of being detected as have found foundation and approach.In state-ownedly carried out genetic test more than 100,000 people, have the working troop more than people more than 100,000, be engaged in the work of genetic test.This present situation can not meet the needs of society far away, will have the development of a large great-leap-forward.Public security and the detection of DNA are medically exactly the one of genetic test.
The numerous gene of field of biological genes and disease, scientific discovery between gene and health and achievement are urgently converted into actual productivity, serve society.Enterprise participation scientific and technical innovation, unites with scientific and technological circle, realizes the conversion of scientific and technological achievement, and the common development for national economy is contributed, and has become the common recognition of the whole society.Verify and improve laboratory scientific payoffs, and being the in-depth research of China's life science, gathering mass data storehouse and large group Sample Storehouse, build new technology platform and support system.
Given this, this patent will utilize SrTiO 3/ CoO-WO 3the signal amplification of nano composite material, the DNA target fragment of the Strength Changes of photic electric signal in system to the extracting after cultivating of true biological cell is utilized to detect, and by developed photoelectricity biology sensor further genralrlization in practical application, the method has that cost is low, highly sensitive, specificity is good, detect the advantages such as quick, and preparation process is comparatively simple, largely overcome the drawback that current DNA detection method is confined to pure biological field, effectively expand the scope of DNA detection method.
Summary of the invention
An object of the present invention is to the biospecific compatibility of DNA and SrTiO based on histidine 3/ CoO-WO 3nano composite material, has prepared one and has possessed specificity, overdelicate photoelectricity biology sensor;
Two of object of the present invention is the detections of the DNA this sensor being used for solid tissue extracting.
Technical scheme of the present invention is as follows:
1. a SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor, step is as follows:
(1) the rectangle ITO electro-conductive glass of 1.0 cm × 2.5 cm is cleaned 30 min with acetone, ultrapure water, EtOH Sonicate successively, purity nitrogen dries up, it can be used as working electrode, platinum filament is to electrode, contrast electrode is saturated calomel electrode, speed is swept with 0.1 V/s, scanning 10 ~ 15 circle, potential range is-0.8 ~ 2.0 V, being in the PBS of 7.0 ~ 9.0 containing the L-Histidine of 0.01 ~ 0.05 mol/L, pH, utilize cyclic voltammetric technology, in working electrode surface electropolymerization L-Histidine, form a strata L-Histidine, preserve in vacuum dryer;
(2) SrTiO of painting 20 ~ 50 μ L is dripped 3/ CoO-WO 3the working electrode surface that nano composite material is modified to (1), dries in 4 DEG C of refrigerators;
(3) continue to drip painting 10 ~ 20 μ L, the upstream primer of target dna of 5 ~ 50 μ g/mL or 10 ~ 20 μ L, 10 ~ 20 μ house keeper g/mL DNA upstream primer to working electrode surface, dry in 4 DEG C of refrigerators;
(4) continue to drip painting 10 ~ 20 μ L, the downstream primer of target dna of 5 ~ 50 μ g/mL or 10 ~ 20 μ L, 10 ~ 20 μ house keeper g/mL DNA downstream primer to working electrode surface, dry in 4 DEG C of refrigerators, obtain a kind of SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor.
2. above-mentioned SrTiO 3/ CoO-WO 3the preparation process of nano composite material is as follows:
(1) SrTiO 3the preparation of solid
By 3 ~ 5 g Ti (OH) 4pressed powder, joins the Sr (NO containing 0.047 ~ 0.189 mol/L 3) 2in solution, vacuum drying obtains pressed powder, and gained pressed powder is placed in muffle furnace, rises to 400 ~ 600 DEG C with the heating rate of 5 DEG C/min, roasting 6 ~ 10 h, deionized water rinsing 4 ~ 8 times, filters, vacuum drying, obtained SrTiO 3solid;
Described Ti (OH) 4pressed powder is dissolved in the absolute ethyl alcohol of 100 mL by 8 ~ 12 g tetrabutyl titanates, abundant stirring and dissolving, and dropwise drip ultrapure water 8 ~ 12 mL, centrifugal 5 ~ 15 min of 5000 r/min, get precipitation, vacuum drying, the Ti (OH) of obtained white 4pressed powder;
Described Sr (NO 3) 2solution is by the Sr (NO of 0.1 ~ 0.4 g 3) 2pressed powder and 0.1 ~ 0.4 g KOH pressed powder, be dissolved in the ultrapure water of 10 mL, the Sr (NO of 0.047 ~ 0.189 obtained mol/L 3) 2solution;
(2) SrTiO 3the preparation of/CoO solid
The SrTiO obtained by (1) 3solid 0.3 ~ 0.7 g is dissolved in the Co (NO of 4 ~ 8 mL, 0.1 ~ 0.3 mol/L 3) 2in solution, dipping 18 ~ 36 h, vacuum drying, is placed in muffle furnace by gained solution, with the ramp to 500 DEG C of 2 ~ 5 DEG C/min, is oxidized 0.5 ~ 2 h at 500 DEG C, obtained SrTiO 3/ CoO black solid powder;
(3) SrTiO 3the preparation of/CoO gel
0.4 ~ 1 g SrTiO obtained by (2) 3/ CoO black solid powder is dissolved in 1 ~ 3 ml ultrapure water, shakes 1 ~ 3 h under room temperature, obtained SrTiO 3/ CoO gel;
(4) WO 3the preparation of solid
By 4 ~ 8 g tungsten powders, be placed in crucible, in muffle furnace, rise to 500 DEG C with the speed of 2 DEG C/min, at 500 DEG C, calcine 2 ~ 4 h, cooling, vacuum drying, obtained WO 3solid;
(5) SrTiO 3/ CoO-WO 3the preparation of nano composite material
By 0.3 ~ 0.6 g SrTiO 3/ CoO gel and 0.1 ~ 0.3 g WO 3solid is dissolved in the ultrapure water of 1 ~ 4 mL, shakes 18 ~ 24 h under room temperature, obtained SrTiO 3/ CoO-WO 3nano composite material.
3. the detecting step of target dna and house-keeping gene thereof
(1) use electrochemical workstation to test with three-electrode system, saturated calomel electrode is contrast electrode, and platinum electrode is auxiliary electrode, prepared SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor is working electrode, tests in the ascorbic acid buffer solution of the PBS of 10 ~ 50 mL, pH 7 ~ 9,0.1 ~ 0.3 mmol/L;
(2) used time m-current method detects target dna standard solution or house keeper DNA standard solution, input voltage is 0.1 V, sample interval 20 s, sample time 20 s, working time 400 s, light source selects 365 nm, 385 nm, 405 nm, 430 nm, 450 nm, 530 nm, 570 nm, 600 nm, and record current changes, drawing curve;
(3) testing sample is replaced target dna standard solution or house keeper DNA standard solution, measure according to the method for drafting of working curve.
useful achievement of the present invention:
(1) L-Histidine has good thermal stability, bioaffinity, in the present invention, L-Histidine is carried out electropolymerization, modify in ITO electrode surface, provide the method that can keep the biomacromolecules such as bioactive immobilized DNA, the method does not need through pre-service simultaneously, and Electrode selectivity, sensitivity and reappearance are better, and electrode response is fast, record the range of linearity 10 pmol/L ~ 30 nmol/L, detect and be limited to 5.4 pmol/L.
(2) the present invention uses SrTiO 3/ CoO-WO 3nano composite material, as photoelectricity beacon material, utilize and the characteristic of coupling can be with to make SrTiO 3, CoO, WO 3the photoelectric characteristic of three all has and significantly strengthens, and substantially increases the photoelectric properties of sensor of the present invention.
(3) DNA that the present invention detects all extracts from actual tumor tissue, has certain practical value.
embodiment:
For further illustrating, illustrate in conjunction with once embodiment:
embodiment 1a kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor
(1) the rectangle ITO electro-conductive glass of 1.0 cm × 2.5 cm is cleaned 30 min with acetone, ultrapure water, EtOH Sonicate successively, purity nitrogen dries up, it can be used as working electrode, platinum filament is to electrode, contrast electrode is saturated calomel electrode, speed is swept with 0.1 V/s, scanning 10 circle, potential range is-0.8 ~ 2.0 V, being in the PBS of 7.4 containing the L-Histidine of 0.01 mol/L, pH, utilize cyclic voltammetric technology, in working electrode surface electropolymerization L-Histidine, form a strata L-Histidine, preserve in vacuum dryer;
(2) SrTiO of painting 20 μ L is dripped 3/ CoO-WO 3the working electrode surface that nano composite material is modified to (1), dries in 4 DEG C of refrigerators
(3) continue to drip a painting 10 μ L, the upstream primer of target dna of 5 μ g/mL or 10 μ L, 10 μ house keeper g/mL DNA upstream primer to working electrode surface, dry in 4 DEG C of refrigerators;
(4) continue to drip a painting 10 μ L, the downstream primer of target dna of 5 μ g/mL or 10 μ L, 10 μ house keeper g/mL DNA downstream primer to working electrode surface, dry in 4 DEG C of refrigerators, obtain a kind of SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor.
embodiment 2a kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor
(1) the rectangle ITO electro-conductive glass of 1.0 cm × 2.5 cm is cleaned 30 min with acetone, ultrapure water, EtOH Sonicate successively, purity nitrogen dries up, it can be used as working electrode, platinum filament is to electrode, contrast electrode is saturated calomel electrode, speed is swept with 0.1 V/s, scanning 12 circle, potential range is-0.8 ~ 2.0 V, being in the PBS of 8.0 containing the L-Histidine of 0.03 mol/L, pH, utilize cyclic voltammetric technology, in working electrode surface electropolymerization L-Histidine, form a strata L-Histidine, preserve in vacuum dryer;
(2) SrTiO of painting 30 μ L is dripped 3/ CoO-WO 3the working electrode surface that nano composite material is modified to (1), dries in 4 DEG C of refrigerators;
(3) continue to drip a painting 15 μ L, the upstream primer of target dna of 25 μ g/mL or 15 μ L, 15 μ house keeper g/mL DNA upstream primer to working electrode surface, dry in 4 DEG C of refrigerators;
(4) continue to drip a painting 15 μ L, the downstream primer of target dna of 25 μ g/mL or 15 μ L, 15 μ house keeper g/mL DNA downstream primer to working electrode surface, dry in 4 DEG C of refrigerators, obtain a kind of SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor.
embodiment 3a kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor
(1) the rectangle ITO electro-conductive glass of 1.0 cm × 2.5 cm is cleaned 30 min with acetone, ultrapure water, EtOH Sonicate successively, purity nitrogen dries up, it can be used as working electrode, platinum filament is to electrode, contrast electrode is saturated calomel electrode, speed is swept with 0.1 V/s, scanning 15 circle, potential range is-0.8 ~ 2.0 V, being in the PBS of 9.0 containing the L-Histidine of 0.05 mol/L, pH, utilize cyclic voltammetric technology, in working electrode surface electropolymerization L-Histidine, form a strata L-Histidine, preserve in vacuum dryer;
(2) SrTiO of painting 50 μ L is dripped 3/ CoO-WO 3the working electrode surface that nano composite material is modified to (1), dries in 4 DEG C of refrigerators;
(3) continue to drip a painting 20 μ L, the upstream primer of target dna of 50 μ g/mL or 20 μ L, 20 μ house keeper g/mL DNA upstream primer to working electrode surface, dry in 4 DEG C of refrigerators;
(4) continue to drip a painting 20 μ L, the downstream primer of target dna of 50 μ g/mL or 20 μ L, 20 μ house keeper g/mL DNA downstream primer to working electrode surface, dry in 4 DEG C of refrigerators, obtain a kind of SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor.
embodiment 4srTiO 3/ CoO-WO 3the preparation of nano composite material
(1) SrTiO 3the preparation of solid
By 3 g Ti (OH) 4pressed powder, joins the Sr (NO containing 0.047mol/L 3) 2in solution, vacuum drying obtains pressed powder, and gained pressed powder is placed in muffle furnace, rises to 400 DEG C with the heating rate of 5 DEG C/min, roasting 6 h, deionized water rinsing 4 times, filters, vacuum drying, obtained SrTiO 3solid;
Described Ti (OH) 4pressed powder is dissolved in the absolute ethyl alcohol of 100 mL by 8 g tetrabutyl titanates, abundant stirring and dissolving, and dropwise drip ultrapure water 8 mL, centrifugal 5 min of 5000 r/min, get precipitation, vacuum drying, the Ti (OH) of obtained white 4pressed powder;
Described Sr (NO 3) 2solution is by the Sr (NO of 0.1 g 3) 2pressed powder and 0.1 KOH pressed powder, be dissolved in the ultrapure water of 10 mL, the Sr (NO of 0.047 obtained mol/L 3) 2solution;
(2) SrTiO 3the preparation of/CoO solid
The SrTiO obtained by (1) 3solid 0.3 g is dissolved in the Co (NO of 4 mL, 0.1 mol/L 3) 2in solution, flood 18 h, vacuum drying, gained solution is placed in muffle furnace, with the ramp to 500 DEG C of 2 DEG C/min, at 500 DEG C, be oxidized 0.5 h, obtained SrTiO 3/ CoO black solid powder;
(3) SrTiO 3the preparation of/CoO gel
The 0.4 g SrTiO obtained by (2) 3/ CoO black solid powder is dissolved in 1 ml ultrapure water, shakes 1 h under room temperature, obtained SrTiO 3/ CoO gel;
(4) WO 3the preparation of solid
By 4 g tungsten powders, be placed in crucible, in muffle furnace, rise to 500 DEG C with the speed of 2 DEG C/min, at 500 DEG C, calcine 2 h, cooling, vacuum drying, obtained WO 3solid;
(5) SrTiO 3/ CoO-WO 3the preparation of nano composite material
By 0.3 g SrTiO 3/ CoO gel and 0.1 g WO 3solid is dissolved in the ultrapure water of 1mL, shakes 18 h under room temperature, obtained SrTiO 3/ CoO-WO 3nano composite material.
embodiment 5srTiO 3/ CoO-WO 3the preparation of nano composite material
(1) SrTiO 3the preparation of solid
By 4 g Ti (OH) 4pressed powder, joins the Sr (NO containing 0.1 mol/L 3) 2in solution, vacuum drying obtains pressed powder, and gained pressed powder is placed in muffle furnace, rises to 500 DEG C with the heating rate of 5 DEG C/min, roasting 8 h, deionized water rinsing 6 times, filters, vacuum drying, obtained SrTiO 3solid;
Described Ti (OH) 4pressed powder is dissolved in the absolute ethyl alcohol of 100 mL by 10 g tetrabutyl titanates, abundant stirring and dissolving, and dropwise drip ultrapure water 10 mL, centrifugal 10 min of 5000 r/min, get precipitation, vacuum drying, the Ti (OH) of obtained white 4pressed powder;
Described Sr (NO 3) 2solution is by the Sr (NO of 0.21 g 3) 2pressed powder and 0.2 g KOH pressed powder, be dissolved in the ultrapure water of 10 mL, the Sr (NO of 0.1 obtained mol/L 3) 2solution;
(2) SrTiO 3the preparation of/CoO solid
The SrTiO obtained by (1) solid 0.5 g is dissolved in the Co (NO of 7 mL, 0.2 mol/L 3) 2in solution, flood 24 h, vacuum drying, gained solution is placed in muffle furnace, with the ramp to 500 DEG C of 3 DEG C/min, at 500 DEG C, be oxidized 1 h, obtained SrTiO 3/ CoO black solid powder;
(3) SrTiO 3the preparation of/CoO gel
The 0.8 g SrTiO obtained by (2) 3/ CoO black solid powder is dissolved in 2 ml ultrapure waters, shakes 2 h under room temperature, obtained SrTiO 3/ CoO gel;
(4) WO 3the preparation of solid
By 6 g tungsten powders, be placed in crucible, in muffle furnace, rise to 500 DEG C with the speed of 2 DEG C/min, at 500 DEG C, calcine 3 h, cooling, vacuum drying, obtained WO 3solid;
(5) SrTiO 3/ CoO-WO 3the preparation of nano composite material
By 0.4 g SrTiO 3/ CoO gel and 0.2 g WO 3solid is dissolved in the ultrapure water of 2 mL, shakes 20 h under room temperature, obtained SrTiO 3/ CoO-WO 3nano composite material.
embodiment 6srTiO 3/ CoO-WO 3the preparation of nano composite material
(1) SrTiO 3the preparation of solid
By 5 g Ti (OH) 4pressed powder, joins the Sr (NO containing 0.189 mol/L 3) 2in solution, vacuum drying obtains pressed powder, and gained pressed powder is placed in muffle furnace, rises to 600 DEG C with the heating rate of 5 DEG C/min, roasting 10 h, deionized water rinsing 8 times, filters, vacuum drying, obtained SrTiO 3solid;
Described Ti (OH) 4pressed powder is dissolved in the absolute ethyl alcohol of 100 mL by 12 g tetrabutyl titanates, abundant stirring and dissolving, and dropwise drip ultrapure water 12 mL, centrifugal 15 min of 5000 r/min, get precipitation, vacuum drying, the Ti (OH) of obtained white 4pressed powder;
Described Sr (NO 3) 2solution is by the Sr (NO of 0.4 g 3) 2pressed powder and 0.4 g KOH pressed powder, be dissolved in the ultrapure water of 10 mL, the Sr (NO of 0.189 obtained mol/L 3) 2solution;
(2) SrTiO 3the preparation of/CoO solid
The SrTiO obtained by (1) 3solid 0.7 g is dissolved in the Co (NO of 8 mL, 0.3 mol/L 3) 2in solution, flood 36 h, vacuum drying, gained solution is placed in muffle furnace, with the ramp to 500 DEG C of 5 DEG C/min, at 500 DEG C, be oxidized 2 h, obtained SrTiO 3/ CoO black solid powder;
(3) SrTiO 3the preparation of/CoO gel
The 1g SrTiO obtained by (2) 3/ CoO black solid powder is dissolved in 3 ml ultrapure waters, shakes 3 h under room temperature, obtained SrTiO 3/ CoO gel;
(4) WO 3the preparation of solid
By 8 g tungsten powders, be placed in crucible, in muffle furnace, rise to 500 DEG C with the speed of 2 DEG C/min, at 500 DEG C, calcine 4 h, cooling, vacuum drying, obtained WO 3solid;
(5) SrTiO 3/ CoO-WO 3the preparation of nano composite material
By 0.6 g SrTiO 3/ CoO gel and 0.3 g WO solid is dissolved in the ultrapure water of 4 mL, shakes 24 h under room temperature, obtained SrTiO 3/ CoO-WO 3nano composite material.
embodiment 7the detection of target dna
(1) use electrochemical workstation to test with three-electrode system, saturated calomel electrode is contrast electrode, and platinum electrode is auxiliary electrode, prepared SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor is working electrode, tests in the ascorbic acid buffer solution of the PBS of 10 mL, pH 7,0.1mmol/L;
(2) m-current method detects target dna standard solution the used time, input voltage is 0.1 V, sample interval 20 s, sample time 20 s, working time 400 s, light source selects 365 nm, 385 nm, 405 nm, 430 nm, 450 nm, 530 nm, 570 nm, 600 nm, and record current changes, drawing curve;
(3) testing sample is replaced target dna standard solution, measure according to the method for drafting of working curve;
(4) range of linearity 10 pmol/L ~ 30 nmol/L, detects and is limited to 5.4 pmol/L.
embodiment 8the detection of house-keeping gene
(1) use electrochemical workstation to test with three-electrode system, saturated calomel electrode is contrast electrode, and platinum electrode is auxiliary electrode, prepared SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor is working electrode, tests in the ascorbic acid buffer solution of the PBS of 50 mL, pH 9,0.3 mmol/L;
(2) used time m-current method detects target dna standard solution or house keeper DNA standard solution, input voltage is 0.1 V, sample interval 20 s, sample time 20 s, working time 400 s, light source selects 365 nm, 385 nm, 405 nm, 430 nm, 450 nm, 530 nm, 570 nm, 600 nm, and record current changes, drawing curve.

Claims (3)

1. a SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor, is characterized in that, comprise the following steps:
(1) the rectangle ITO electro-conductive glass of 1.0 cm × 2.5 cm is cleaned 30 min with acetone, ultrapure water, EtOH Sonicate successively, purity nitrogen dries up, it can be used as working electrode, platinum filament is to electrode, contrast electrode is saturated calomel electrode, speed is swept with 0.1 V/s, scanning 10 ~ 15 circle, potential range is-0.8 ~ 2.0 V, being in the PBS of 7.0 ~ 9.0 containing the L-Histidine of 0.01 ~ 0.05 mol/L, pH, utilize cyclic voltammetric technology, in working electrode surface electropolymerization L-Histidine, form a strata L-Histidine, preserve in vacuum dryer;
(2) SrTiO of painting 20 ~ 50 μ L is dripped 3/ CoO-WO 3the working electrode surface that nano composite material is modified to (1), dries in 4 DEG C of refrigerators;
(3) continue to drip painting 10 ~ 20 μ L, the upstream primer of target dna of 5 ~ 50 μ g/mL or 10 ~ 20 μ L, 10 ~ 20 μ house keeper g/mL DNA upstream primer to working electrode surface, dry in 4 DEG C of refrigerators;
(4) continue to drip painting 10 ~ 20 μ L, the downstream primer of target dna of 5 ~ 50 μ g/mL or 10 ~ 20 μ L, 10 ~ 20 μ house keeper g/mL DNA downstream primer to working electrode surface, dry in 4 DEG C of refrigerators, obtain a kind of SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor.
2. a kind of SrTiO as claimed in claim 1 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor, wherein, described SrTiO 3/ CoO-WO 3the preparation process of nano composite material is as follows:
(1) SrTiO 3the preparation of solid
By 3 ~ 5 g Ti (OH) 4pressed powder, joins the Sr (NO containing 0.047 ~ 0.189 mol/L 3) 2in solution, vacuum drying obtains pressed powder, and gained pressed powder is placed in muffle furnace, rises to 400 ~ 600 DEG C with the heating rate of 5 DEG C/min, roasting 6 ~ 10 h, deionized water rinsing 4 ~ 8 times, filters, vacuum drying, obtained SrTiO 3solid;
Described Ti (OH) 4pressed powder is dissolved in the absolute ethyl alcohol of 100 mL by 8 ~ 12 g tetrabutyl titanates, abundant stirring and dissolving, and dropwise drip ultrapure water 8 ~ 12 mL, the centrifugal 5 ~ 15min of 5000 r/min, gets precipitation, vacuum drying, the Ti (OH) of obtained white 4pressed powder;
Described Sr (NO 3) 2solution is by the Sr (NO of 0.1 ~ 0.4 g 3) 2pressed powder and 0.1 ~ 0.4 g KOH pressed powder, be dissolved in the ultrapure water of 10 mL, the Sr (NO of 0.047 ~ 0.189 obtained mol/L 3) 2solution;
(2) SrTiO 3/ CoO 3the preparation of solid
The SrTiO obtained by (1) 3solid 0.3 ~ 0.7 g is dissolved in the Co (NO of 4 ~ 8 mL, 0.1 ~ 0.3 mol/L 3) 2in solution, dipping 18 ~ 36 h, vacuum drying, is placed in muffle furnace by gained solution, with the ramp to 500 DEG C of 2 ~ 5 DEG C/min, is oxidized 0.5 ~ 2 h at 500 DEG C, obtained SrTiO 3/ CoO black solid powder;
(3) SrTiO 3the preparation of/CoO gel
0.4 ~ 1 g SrTiO obtained by (2) 3/ CoO black solid powder is dissolved in 1 ~ 3 ml ultrapure water, shakes 1 ~ 3 h under room temperature, obtained SrTiO 3/ CoO gel;
(4) WO 3the preparation of solid
By 4 ~ 8 g tungsten powders, be placed in crucible, in muffle furnace, rise to 500 DEG C with the speed of 2 DEG C/min, at 500 DEG C, calcine 2 ~ 4 h, cooling, vacuum drying, obtained WO 3solid;
(5) SrTiO 3/ CoO-WO 3the preparation of nano composite material
By 0.3 ~ 0.6 g SrTiO 3/ CoO gel and 0.1 ~ 0.3 g WO 3solid is dissolved in the ultrapure water of 1 ~ 4 mL, shakes 18 ~ 24 h under room temperature, obtained SrTiO 3/ CoO-WO 3nano composite material.
3. a kind of SrTiO of preparing of preparation method as claimed in claim 1 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor is used for the detection of target dna and house-keeping gene thereof, it is characterized in that:
(1) use electrochemical workstation to test with three-electrode system, saturated calomel electrode is contrast electrode, and platinum electrode is auxiliary electrode, prepared SrTiO 3/ CoO-WO 3nano composite material photoelectricity aptamer sensor is working electrode, tests in the ascorbic acid buffer solution of the PBS of 10 ~ 50 mL, pH 7 ~ 9,0.1 ~ 0.3 mmol/L;
(2) used time m-current method detects target dna standard solution or house keeper DNA standard solution, input voltage is 0.1 V, sample interval 20 s, sample time 20 s, working time 400 s, light source selects 365 nm, 385 nm, 405 nm, 430 nm, 450 nm, 530 nm, 570 nm, 600 nm, and record current changes, drawing curve;
(3) testing sample is replaced target dna standard solution or house keeper DNA standard solution, measure according to the method for drafting of working curve.
CN201410601204.0A 2014-11-01 2014-11-01 A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application Expired - Fee Related CN104330454B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410601204.0A CN104330454B (en) 2014-11-01 2014-11-01 A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201410601204.0A CN104330454B (en) 2014-11-01 2014-11-01 A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application

Publications (2)

Publication Number Publication Date
CN104330454A CN104330454A (en) 2015-02-04
CN104330454B true CN104330454B (en) 2015-10-21

Family

ID=52405222

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410601204.0A Expired - Fee Related CN104330454B (en) 2014-11-01 2014-11-01 A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application

Country Status (1)

Country Link
CN (1) CN104330454B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106390986B (en) * 2016-11-02 2019-05-07 西北师范大学 A kind of preparation method of pucherite/strontium titanates composite photo-catalyst

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007102629A1 (en) * 2005-06-28 2007-09-13 Korea Research Institute Of Chemical Technology Carbon nanotube transistor biosensors with aptamers as molecular recognition elements and method for sensing a target material using the same
CN102495125A (en) * 2011-12-07 2012-06-13 北京航空航天大学 Method for detecting L-histidine by use of nano-gold crystal of DNA (deoxyribonucleic acid) sequence self-breaking and high-energy index surface
CN103940867A (en) * 2014-04-11 2014-07-23 同济大学 Method for preparing photoelectric adapter sensor for detecting 17beta-estradiol

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005069692A (en) * 2003-08-22 2005-03-17 Tsutomu Nagaoka Amino acid analyzer

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007102629A1 (en) * 2005-06-28 2007-09-13 Korea Research Institute Of Chemical Technology Carbon nanotube transistor biosensors with aptamers as molecular recognition elements and method for sensing a target material using the same
CN102495125A (en) * 2011-12-07 2012-06-13 北京航空航天大学 Method for detecting L-histidine by use of nano-gold crystal of DNA (deoxyribonucleic acid) sequence self-breaking and high-energy index surface
CN103940867A (en) * 2014-04-11 2014-07-23 同济大学 Method for preparing photoelectric adapter sensor for detecting 17beta-estradiol

Also Published As

Publication number Publication date
CN104330454A (en) 2015-02-04

Similar Documents

Publication Publication Date Title
CN104297323B (en) The carboxylated C of a kind of ZnO CdTe- 3n 4the preparation of photoelectricity DNA sensor and application thereof
CN105004775B (en) Curing object point/nanometer sheet compound DNA electrochemical probe and its preparation method and application
CN104316460B (en) A kind of TiO 2the preparation method of-CdSe nano composite material photoelectricity biology sensor and application thereof
CN112461905B (en) Construction method of novel photo-assisted bipolar self-powered adapter sensor
CN102854235B (en) Preparation method and application of gynecological tumor marker immune sensor constructed with ordered mesoporous carbon
CN104833806B (en) A kind of preparation method of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor and application
Shen et al. Ultrasensitive aptasensor for isolation and detection of circulating tumor cells based on CeO2@ Ir nanorods and DNA walker
CN103743802A (en) DNA biosensor based on three-dimensional ordered gold doped nano titanium dioxide electrode as well as preparation method and application of DNA biosensor
CN104931324A (en) Hematoxylin and eosin staining kit for circulating tumor cells (CTC)
CN103940867A (en) Method for preparing photoelectric adapter sensor for detecting 17beta-estradiol
CN103134793A (en) Electrogenerated chemiluminescence sensor with high sensitivity in cancer cell detection and fabrication method of sensor
CN104330454B (en) A kind of SrTiO 3/ CoO-WO 3the preparation method of nano composite material photoelectricity aptamer sensor and application
CN108690870A (en) The preparation method of biosensor is quenched in tris (bipyridine) ruthenium electrochemical luminescence
CN108088882B (en) Electrochemical detection method of stem cells
CN109298038A (en) A kind of MnCo2O4The preparation method and application of/Polyaniline-modified Glassy Carbon Electrode
CN105911128A (en) Enzyme-free glucose electrochemical sensor and application thereof
CN105158125B (en) A kind of telomere length measuring method
CN104316705B (en) A kind of hybridization indicator 5, preparation method and the purposes of 7-dinitro-2-sulfo group-acridone
CN103454266A (en) Method for preparing cadmium sulfide-coated zinc oxide nano array composite material and method for detecting multiple tumor cells
CN108508068B (en) Method for detecting HER2 gene specific sequence by anion porphyrin-carbon nanotube modified electrode
CN105784808B (en) A kind of TiO2‑CH3NH3PbI3The application of photoelectricity M2 macrophage sensors
CN105675679A (en) Preparation and application of ZnO-NCQDs (Nitrogen-doped Carbon Quantum Dots) DNA photoelectric sensor
CN110907416A (en) Circulating tumor cell detection device based on hollow nano needle tube electroporation system and detection method thereof
CN105758912B (en) A kind of nano-TiO2‑MoS2The application of the cell cell sensors of photoelectricity Saos 2
CN102586412B (en) MicroRNA (ribonucleic acid) specific expression profile and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20151021

Termination date: 20191101