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

A rapid detection method for microRNA-7a based on nano-gold colorimetry

A detection method, nano-gold technology, applied in the fields of molecular biology and nucleic acid chemistry, can solve the problems of destruction, cumbersome biological sample activity, high instrument cost, etc., and achieve the effect of mild reaction environment and pollution-free, short detection cycle and fast analysis speed

Active Publication Date: 2021-07-13
GUILIN UNIVERSITY OF TECHNOLOGY
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One is the labeling method, which has high sensitivity, but involves synthesis, purification, labeling and other processes in the labeling process, which is too cumbersome and will damage the activity of biological samples; the other is the label-free method, but its instrument cost is high, Requires more professional operating techniques, making its application subject to greater limitations

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
  • A rapid detection method for microRNA-7a based on nano-gold colorimetry
  • A rapid detection method for microRNA-7a based on nano-gold colorimetry
  • A rapid detection method for microRNA-7a based on nano-gold colorimetry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Preparation of gold nanoparticles: Pipette 100 mL of chloroauric acid solution with a concentration of 0.01% by mass and pour it into a round-bottomed flask, place it in an oil bath, heat it to boil, and quickly add 1.5 mL with a concentration of 1% by mass % sodium citrate solution as a reducing agent, continue heating for 15 minutes, observe that the color of the solution gradually changes from initial light yellow to wine red, and finally stop heating and cool to room temperature, and the whole process is carried out under stirring. Gold nanoparticles with a diameter of about 24nm were obtained.

[0030] (2) Design the probe Probe (base number 34mer) complementary to the target miRNA-7a.

[0031] (3) After mixing the target miRNA-7a with a concentration of 131.57 nmol / L and the probe Probe with a substance amount of 80 pmol and a base number of 34mer, place it at 38°C for 30 minutes and hybridize to obtain a hybridization mixed solution.

[0032] (4) Put the mix...

Embodiment 2

[0038] (1) Preparation of gold nanoparticles: Pipette 100 mL of chloroauric acid solution with a concentration of 0.01% by mass and pour it into a round-bottomed flask, place it in an oil bath, heat it to boil, and quickly add 1.5 mL with a concentration of 1% by mass % sodium citrate solution as reducing agent, continue to heat for 15 minutes, observe that the color of the solution gradually changes from initial light yellow to wine red, and finally stop heating and cool to room temperature, and carry out under stirring throughout the whole process; the obtained particle size is approximately 24nm gold nanoparticles.

[0039] (2) Design probes complementary to the target miRNA-7a, with base numbers of 22mer, 34mer, and 40mer.

[0040](3) After mixing the target miRNA-7a with a concentration of 131.57 nmol / L and 80 pmol of substances, probes with base numbers of 22mer, 34mer, and 40mer were mixed, and hybridized at 38°C for 30 minutes to obtain hybridization mixture.

[0041...

Embodiment 3

[0047] (1) Preparation of gold nanoparticles: Pipette 100 mL of chloroauric acid solution with a concentration of 0.01% by mass and pour it into a round-bottomed flask, place it in an oil bath, heat it to boil, and quickly add 1.5 mL with a concentration of 1% by mass % sodium citrate solution as reducing agent, continue to heat for 15 minutes, observe that the color of the solution gradually changes from initial light yellow to wine red, and finally stop heating and cool to room temperature, and carry out under stirring throughout the whole process; the obtained particle size is approximately 24nm gold nanoparticles.

[0048] (2) Design a 34mer probe complementary to the target miRNA-7a.

[0049] (3) After mixing the target miRNA-7a with a concentration of 131.57 nmol / L and the probe probe with a concentration of 40, 80, 120, 160, and 200 pmol and a base number of 34mer, place it at 38°C for 30 Minutes, the hybridization mixture solution was obtained.

[0050] (4) Put the m...

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

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a rapid detection method for microRNA-7a based on nano-gold colorimetry. Using the electrostatic difference between single-stranded DNA and double-stranded DNA, the different interactions (color changes) with gold nanometers under high salt concentration are used for colorimetric detection of the target, and the target is indirectly detected by combining the absorbance value of the UV-visible spectrometer. concentration. The detection concentration lower limit of the present invention reaches 5.3nmol / L, and the linear regression equation is A=0.002C+0.238, r=0.990. This method can not only perform specific analysis on unlabeled microRNA-7a, but also distinguish single-base mismatch sequences well, with strong specificity, simple method, fast analysis speed, and the lower limit of detection of the concentration of the target visible to the naked eye. It can be widely used in practical detection.

Description

technical field [0001] The invention relates to a rapid detection method for microRNA-7a based on nano-gold colorimetry, belonging to the fields of molecular biology and nucleic acid chemistry. Background technique [0002] MicroRNA (miRNA), as a very important non-coding protein RNA, is about 19-25 bases in length and widely exists in plants, viruses, and mammals. It plays an important regulatory function in cell proliferation, cell death, tumorigenesis, and mammalian cell growth. The short miRNA sequence has high similarity and low content. Therefore, it is necessary to develop a sensitive and selective miRNA detection method. Early diagnosis provides new ideas. [0003] Currently, there are two main methods for miRNA detection. One is the labeling method, which has high sensitivity, but involves synthesis, purification, labeling and other processes in the labeling process, which is too cumbersome and will damage the activity of biological samples; the other is the label...

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
Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N21/78C12Q1/6825
CPCC12Q1/68G01N21/31G01N21/78
Inventor 朱文远梁晓琳周琳莹李丹温其霖潘宏程
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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