Supercharge Your Innovation With Domain-Expert AI Agents!

Nucleic acid detection method combining DNA/Fe3O4 reticular structure with magnetic three-phase extraction method

A network structure and detection method technology, applied in the field of nucleic acid detection, can solve the problems of high detection limit and achieve the effects of lower detection limit, simple operation and high sensitivity

Pending Publication Date: 2021-02-12
JIANGSU UNIV OF SCI & TECH
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the detection relies only on the hybridization of DNA, the detection limit is too high, usually in the pM to nM range, thus limiting its practical application

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
  • Nucleic acid detection method combining DNA/Fe3O4 reticular structure with magnetic three-phase extraction method
  • Nucleic acid detection method combining DNA/Fe3O4 reticular structure with magnetic three-phase extraction method
  • Nucleic acid detection method combining DNA/Fe3O4 reticular structure with magnetic three-phase extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) 1.8g FeCl 3 ·6H 2 O and 0.9 g FeSO 4 ·7H 2 O was dissolved in a beaker with 50 mL of pure water, then transferred to a three-neck round bottom flask, stirred magnetically, and washed with N 2 Purge the mixture, and when the reaction temperature rises to 80°C, slowly add 10 mL of 25% NH 4 OH, the solution turned black rapidly. The reaction mixture was stirred in a water bath at 80°C for 30 min, and continuously heated with N 2 purge and then separate the Fe with a magnet 3 o 4 Nanosheets and supernatant, and remove the supernatant, wash with boiled ultrapure water 5 times. Redissolve the nanomaterials in 15 mL of ultrapure water, add 1.25 g of citric acid, and stir at 90 °C for 2 h, then dialyze in ultrapure water for 24 h with a 12KDa dialysis membrane, in which the water is changed every 8 h and cooled to room temperature. Then, Fe 3 o 4 The nanosheets were washed by centrifugation at 1000rpm for 10min, and the Fe 3 o 4The nanosheet solution was centrif...

Embodiment 2

[0050] The difference between this embodiment and embodiment 1 is that in step 3, it is an animal serum sample containing DNA, and the sequences used include:

[0051]

[0052] Table 2 is the table of detection results of animal serum samples with different DNA contents, from which it can be seen that the relative recoveries when different contents of DNA are mixed into the samples are all close to 100%, indicating that the method can accurately quantify the DNA in the samples.

[0053] Table 2 Detection results of samples with different DNA contents

[0054] amount of DNA incorporated relative recovery Sample 6 0 - Sample 7 0.6fM 100.7 Sample 8 6f 104.9 Sample 9 60fM 101.2 sample 10 600fM 103.8

[0055] Relative recovery = (total concentration - blank concentration) / spiked concentration

[0056] It can be seen from Figure 4(a) and (b) that there is a linear positive correlation between the concentration of DNA 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 a nucleic acid detection method combining a DNA / Fe3O4 reticular structure with a magnetic three-phase extraction method. The nucleic acid detection method comprises the following steps: (1) preparing Fe3O4 nanosheets; (2) modifying single-stranded DNA on the Fe3O4 nanosheets to obtain Fe3O4 / ssDNA; (3) preparing a hyperbranched DNA structure sample, and mixing the hyperbranched DNA structure sample with Fe3O4 / ssDNA for reaction to obtain a DNA / Fe3O4 reticular structure; and (4) taking the system of the DNA / Fe3O4 reticular structure as a water phase, mixing with an organic phase, immersing a magnetic rod with extraction liquid drops into the organic phase, extracting the DNA / Fe3O4 reticular structure, taking out, monitoring the liquid drops subjected to the catalyticchromogenic reaction by using an ultraviolet-visible spectrograph, and analyzing ultraviolet-visible spectral data to obtain the content of the target nucleic acid in a sample to be detected. According to the method, the detection limit of nucleic acid is greatly reduced, the practicability is high, the detection limit of microRNA-122 is as low as 0.147 aM, the detection limit of H-DNA is as low as 0.34 aM, the linear ranges are respectively 0.5 aM to 1 pM and 1 aM to 1 pM (r2 is greater than 0.995), and a single-drop microextraction technology is utilized, so that the detection efficiency ishigh, the operation is simple and the cost is low.

Description

technical field [0001] The present invention relates to a nucleic acid detection method, more specifically, to a DNA / Fe 3 o 4 Nucleic acid detection method combined with network structure and magnetic three-phase extraction method. Background technique [0002] The components in biological fluids are complex, so the detection of certain nucleic acids (microRNA or DNA) is easily affected by the matrix effect, resulting in the inaccuracy of existing detection methods not being high enough. Most current nucleic acid detection methods rely on hybridization, such as the hybridization of target microRNA or DNA molecules with complementary labeled oligonucleotide probes. However, nucleic acid hybridization alone cannot meet the sensitivity and accuracy requirements. In recent years, the development of nanotechnology combining DNA with nanomaterials provides a new signal amplification strategy for biosensors for early diagnosis. DNA nanotechnology can not only use the rigid stra...

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): C12Q1/682
CPCC12Q1/682C12Q2523/308C12Q2563/159C12Q2563/125C12Q2525/207
Inventor 唐盛祁桐姚瑶唐良秀陈文慧沈薇鞠嘉和
Owner JIANGSU UNIV OF SCI & TECH
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com