Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Amperometric DNA (deoxyribonucleic acid) electrochemical sensor based on protein controlled assembling interface

A technology for controlling assembly and protein, applied in the field of biosensing, can solve problems affecting sensor performance, difficult to control probe density and distribution, DNA probe assembly density affects interaction, etc., and achieve good reproducible results

Inactive Publication Date: 2012-07-04
FUJIAN MEDICAL UNIV
View PDF5 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The assembly density of DNA probes largely affects the interaction between DNA probes and complementary strands, which in turn affects the performance of sensors.
At present, DNA electrochemical sensors generally adopt the method of direct self-assembly, that is, to modify the groups at the end of the probe DNA, and through the chemical bonding between the modified groups and the substrate electrode, the probe DNA is fixed to the electrode surface. Very random, it is difficult to control the density and distribution of probes, thus causing the randomness of the hybridization reaction, which is one of the main reasons for the poor reproducibility of the sensor

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Amperometric DNA (deoxyribonucleic acid) electrochemical sensor based on protein controlled assembling interface
  • Amperometric DNA (deoxyribonucleic acid) electrochemical sensor based on protein controlled assembling interface
  • Amperometric DNA (deoxyribonucleic acid) electrochemical sensor based on protein controlled assembling interface

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The preparation steps of the amperometric DNA electrochemical sensor based on the protein assembly interface are as follows:

[0048] (1) Piranha solution (30 wt% H 2 o 2 Concentrated H with a concentration of 98wt% 2 SO 4 , mixed at a volume ratio of 1:3) sonicated for 10 min, ultrasonically cleaned with deionized water twice for 5 min each, and then washed with 0.3 μm and 0.05 μm Al 2 o 3 The mixture of water and water is polished to a mirror surface, followed by ultrasonic cleaning with ethanol and distilled water. Place the sonicated electrode at 0.5 M H 2 SO 4 In the 0~1.6 V potential range, the cyclic voltammetry scan was stable, washed with double distilled water, N 2 Blow dry for later use;

[0049] (2) Submerge the bare gold electrode treated in step (1) in 0.5 wt % BSA solution, assemble at room temperature for 15 min, wash with PBS buffer solution of pH 7.40 and double distilled water, and blow dry with nitrogen. Take 4 μl capture probe DNA (1 μmol / L...

Embodiment 2

[0051] The detection steps of the target DNA by the amperometric DNA electrochemical sensor based on the protein assembly interface are as follows:

[0052] (1) The capture probe DNA (5'-SH-(CH 2 ) 6 -T 10 -CTTCA GAACT GCTGC TCTGG GTCTC AATGG - 3') with fully complementary DNA sequence (5'-ACCAC GTGGC CAGTG GCGCC GGGGA GGCAG CCATT GAGAC CCAGA GCAGC AGTTC TGAAG - 3') and signal probe DNA (5'-CTGCC TCCCC GGCGC CACTG GCCAC GTGGT-Biotin-3') (oligonucleotide, synthesized by Baosheng Bioengineering Co., Ltd.) in the hybridization buffer solution (made of 10 mmol / L Na 2 HPO 4 -NaH 2 PO 4 and 1 mol / L NaCl, and with H 3 PO 4 Adjust the pH to 7.40 with NaOH, and hybridize in a 50°C water bath for 35 min to form double-stranded DNA, then wash with PBS buffer at pH 7.40, double-distilled water, and blow dry with nitrogen. Add 3 μl of HRP-avidin solution (0.5 U / mL) dropwise on the surface of the electrode, react at room temperature for 15 min, stir and wash with PBS washing solutio...

Embodiment 3

[0056] The detection steps of the target DNA by the amperometric DNA electrochemical sensor based on the protein assembly interface are as follows:

[0057] (1) The capture probe DNA (5'-SH-(CH 2 ) 6 -T 10 -CTTCA GAACT GCTGC TCTGG GTCTC AATGG - 3') with a single base mismatch DNA sequence (5'-ACCAC GTGGC CAGTC GCGCC GGGGA GGCAG CCATT GAGAC CCAGA GCAGC AGTTC TGAAG - 3') and signal probe DNA (5'-CTGCC TCCCC GGCGC CACTG GCCAC GTGGT-Biotin-3') (oligonucleotide, synthesized by Baosheng Bioengineering Co., Ltd.) in hybridization buffer solution (made of 10 mmol / L Na 2 HPO 4 -NaH 2 PO 4 and 1 mol / L NaCl, and with H 3 PO 4 The pH was adjusted to 7.40 with NaOH, and the experimental water was double-distilled water) for 35 min in a water bath at 50 °C, followed by washing with PBS buffer at pH 7.40, double-distilled water, and drying with nitrogen. Add 3 μl of HRP-avidin solution (0.5 U / mL) dropwise on the surface of the electrode, react at room temperature for 15 min, stir and...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an amperometric DNA (deoxyribonucleic acid) electrochemical sensor based on a protein controlled assembling interface. The amperometric DNA electrochemical sensor comprises an electrode, proteins, a capture probe DNA, a signal probe DNA and horse radish peroxidases, wherein the electrode is a gold electrode, the proteins are bovine serum albumins, the capture probe DNA is a 5' terminated mercapto modified single-strand DNA, the signal probe DNA is a 3' terminated biotin labeled single-strand DNA, and the horse radish peroxidases are labeled by avidin. The amperometric DNA electrochemical sensor is characterized in that the bovine serum albumins are formed into a sequentially-assembled bovine serum albumin molecule layer on the surface of the gold electrode, and the capture probe DNA is assembled by using sequential holes among bovine serum albumin molecules in the bovine serum albumin molecule layer. The amperometric DNA electrochemical sensor is used for carrying out detection on a target DNA, and the detection limit is 0.01pmol / L. Compared with similar methods, the amperometric DNA electrochemical sensor constructed by using the method is high in sensitivity and good in repeatability.

Description

technical field [0001] The invention relates to a preparation method of an ampere-type DNA electrochemical sensor based on a protein-controlled assembly interface, and belongs to the technical field of biosensing. Background technique [0002] At present, the amperometric DNA electrochemical sensor constructed by the combination of electrochemical and enzyme catalysis technology has received extensive attention, and has been applied to the research of gene detection related to diseases such as tumors. The basic principle is: use single-stranded DNA as a capture probe, modify the sulfhydryl group at its end, and fix it to the surface of the gold electrode by low-temperature self-assembly through the combination of sulfhydryl group and gold. This capture probe chain can be combined with one end of the target DNA. Hybridization, the other end of the captured target DNA hybridizes with the signal probe strand to form a "sandwich" structure. The 3'-end of the signal probe is lab...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N27/26G01N27/327
Inventor 陈伟刘银环林新华刘爱林陈元仲
Owner FUJIAN MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com