Method for detecting nucleotide sequence by using poly(methyl acrylic pyrene-poly (methyl) acrylic dimethylamine ethyl ester copolymer, and product thereof

A technology of dimethylamine ethyl acrylate and polypyrene methyl acrylate, which is applied in the field of functional polymers, can solve the problem of reducing the thermal stability of the hybrid double-chain of fluorescent probes and targeting molecules, complicated preparation of labeled fluorescent probes, Reduce the detection accuracy of fluorescent probes and other problems, and achieve the effect of simple and easy synthesis route, reduced fluorescence intensity, good selectivity and sensitivity

Inactive Publication Date: 2013-02-20
UNIV OF SCI & TECH BEIJING
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research results show that the disadvantage of labeled fluorescent probes for detecting nucleic acid molecules is that the introduction of fluorescent groups will reduce the thermal stability of the hybrid double strands of fluorescent probes and target molecules, thereby reducing the detection accuracy of fluorescent probes ; Labeled fluorescent probes need to link fluorescent groups to oligonucleotide molecular chains through covalent bonds to achieve specific recognition of nucleic acid molecules, and the synthesis and operation are more complicated
This patent has invented a new non-labeled fluorescent polyelectrolyte, which is electrostatically interacted with DNA to form a new type of non-labeled fluorescent probe. This fluorescent polymer overcomes the non-specific recognition of small molecule fluorescent probes And the preparation of labeled fluorescent probes is complicated and other shortcomings, it opens up a new test method for the detection of biological macromolecules

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
  • Method for detecting nucleotide sequence by using poly(methyl acrylic pyrene-poly (methyl) acrylic dimethylamine ethyl ester copolymer, and product thereof
  • Method for detecting nucleotide sequence by using poly(methyl acrylic pyrene-poly (methyl) acrylic dimethylamine ethyl ester copolymer, and product thereof
  • Method for detecting nucleotide sequence by using poly(methyl acrylic pyrene-poly (methyl) acrylic dimethylamine ethyl ester copolymer, and product thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0033] Add 2ml of dry THF solvent to a 25ml clean single-necked round bottom flask, then add 0.3mmol copolymer and 0.9mmol methyl iodide respectively, and react at room temperature for 24 hours. After the reaction, filter the solvent THF, and then Extract the product with a Soxhlet extractor, wherein the extractant is tetrahydrofuran, and the extraction time is 12 hours, and finally the final fluorescent polyelectrolyte (P(DMAEMA + -co-Py)) was dried in a vacuum oven at 70°C for 12 hours.

example 2

[0035] (1) Sample preparation: Weigh an appropriate amount of fluorescent polyelectrolyte P(DMAEMA + -co-Py) was dissolved in buffer PBS to make its original concentration 1×10 -5 M, use 1ml of buffer to dilute each tube containing 1od of DNA1, DNA2, DNA3, DNAa, DNAc, DNAt, wherein DNA2 is a hairpin structure, DNA1, DNA3, DNAa, DNAc and DNAt are linear structures, and DNA2 is partially complementary to DNA1, and completely non-complementary to DNAa, DNAc, and DNAt; DNA1 is completely complementary to DNA3, and completely non-complementary to DNAa, DNAc, and DNAt. Then take appropriate amount of fluorescent polyelectrolyte buffer and DNA buffer, and finally make P(DMAEMA + -co-Py), P(DMAEMA + -co-Py)+DNA1,P(DMAEMA + -co-Py)+DNA1+DNA3, P(DMAEMA + -co-Py)+DNA1+DNAa, P(DMAEMA + -co-Py)+DNA1+DNAc, P(DMAEMA + -co-Py)+DNA1+DNAt, P(DMAEMA + -co-Py)+DNA2,P(DMAEMA + -co-Py)+DNA2+DNA1, P(DMAEMA + -co-Py)+DNA2+DNAa, P(DMAEMA + -co-Py)+DNA2+DNAc, P(DMAEMA + -co-Py)+DNA2+DNAt11 s...

example 3

[0039] (1) Sample preparation: Weigh an appropriate amount of fluorescent polyelectrolyte P(DMAEMA + -co-Py) was dissolved in buffer PBS to make its original concentration 1×10 -5 M, use 1ml of buffer to dilute DNA1, DNA2, and DNA3 containing 1od in each tube, where DNA2 has a hairpin structure, DNA1, and DNA3 are linear structures, and DNA2 is partially complementary to DNA1, and DNA1 is completely complementary to DNA3. Then take appropriate amount of fluorescent polyelectrolyte buffer and DNA buffer, and finally make P(DMAEMA + -co-Py)+DNA1+DNA3, DNA1+DNA3, P(DMAEMA + -co-Py)+DNA2+DNA1, DNA2+DNA14 sample solutions, wherein in each sample solution, the concentration of fluorescent polyelectrolyte remains 4.0×10 -6 M, the concentration of DNA was kept at 1×10 -4 M.

[0040] (2) Sample heat treatment: Put the sample prepared in (1) into a 90°C oven for heat treatment for 5 minutes, and then quickly move to a 40°C oven for half an hour.

[0041] Finally, circular dichroism...

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 relates to a method for detecting nucleotide sequence by using poly(methyl acrylic pyrene-poly (methyl) acrylic dimethylamine ethyl ester copolymer, and a product thereof, and belongs to the field of functional polymers. The method comprises the steps of protonating the poly(methyl acrylic pyrene-poly (methyl) acrylic dimethylamine ethyl ester copolymer with alkyl halide to obtain a polyelectrolyte having positive charges, and acting the polyelectrolyte with a linear or a hairpin structure DNA to form a novel fluorescent probe for detecting DNA. The method is advantageous in that the product is the novel fluorescent probe which is simple in preparation, easy to operate and effective, provides a novel approach for stable, efficient and specific identification of the nucleic acid molecules, and has significant scientific and application value in aspects of disclosing diseases and genetic variations and the like.

Description

technical field [0001] A method for detecting nucleic acid sequences using polypyrene methyl acrylate-poly(meth)dimethylaminoethyl acrylate copolymers, in particular to a method of using non-conjugated fluorescent copolymers and straight-chain or hairpin-structured DNA through electrostatic The method for forming a novel molecular probe to detect target DNA belongs to the technical field of functional macromolecules. Background technique [0002] The application of functional polymers in the field of biomedicine, especially the study of fluorescent polymers in the analysis and detection of biological macromolecules, has attracted close attention of scientists at home and abroad, and this research direction is also a research hotspot in frontier science in the world. Some small molecular fluorescent dyes such as acridine, phenanthridine dyes, cyanine dyes, fluorescein and rhodamine dyes, thiazine and oxazine dyes are currently widely used nucleic acid fluorescent probes. Thes...

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 Applications(China)
IPC IPC(8): C12Q1/68G01N21/64C08F220/34C08F220/18C08F8/44
Inventor 王国杰杨领叶赵敏董杰张瑞辰
Owner UNIV OF SCI & TECH BEIJING
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap