Solid phase nucleic acid detection probe and preparing method thereof

A technology for detecting probes and oligonucleotide probes, which is used in biochemical equipment and methods, microbial determination/inspection, fluorescence/phosphorescence, etc. The problems of mutation detection and single base mismatch detection are difficult to achieve the effect of improving the mismatch recognition ability, shortening the diagnosis time, and reducing the detection cost.

Inactive Publication Date: 2005-06-15
SOUTHEAST UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this process, the reliability of the detection is affected due to the problem of labeling efficiency.
2) Although there are currently some technologies for non-labeled detection of gene sequences, they are not very mature
3) Single base mismatch detection is still a difficult problem at present. The main method is to determine the sequence of nucleic acid, which is not accurate and convenient enough for the determination of heterozygous gene mutations.
(4) The detection method of primer specificity cannot be used for the detection of heterozygous gene mutation

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
  • Solid phase nucleic acid detection probe and preparing method thereof
  • Solid phase nucleic acid detection probe and preparing method thereof
  • Solid phase nucleic acid detection probe and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0022] 4. Preparation of solid-phase nucleic acid probes: a commercial solid-phase chemical synthesis method was used to synthesize the equipment

[0023] A well-planned oligonucleotide probe comprising at least one fluorescent chromophore-specific nucleic acid sequence; the method for preparing the nucleic acid probe can also be to directly synthesize nucleic acid on a fluorescence quenching material by using an in-situ synthesis method and fluorescent labeling.

[0024] 5. Immobilization of probes: The probes synthesized by solid-phase chemistry are transferred to the surface of the solid substrate by machines or other means, and connected to the solid substrate under appropriate conditions. Each dot contains at least two probes labeled with different fluorophores.

[0025] 6. Hybridization and detection: Appropriate ions and buffers are added to the tested system, and the target gene and the solid-phase probe of the present invention are subjected to hybridization reaction, ...

Embodiment 1

[0026] Example 1, in situ synthetic non-marker gene chip

[0027] 1. Cleaning of glass slides: Soak the slides in lotion overnight, rinse, then soak in alkali ethanol solution for two hours, rinse with double distilled water, and blow dry with nitrogen for later use.

[0028] 2. Glass slide modification: Take clean slides, soak them in the acetone solution of triethoxyaminosilane for 5 minutes, wash them, bake them at 100 degrees for 40 minutes, soak them in glutaraldehyde solution for 2 hours, and wash them with nitrogen. blow dry.

[0029] 3. Gold nanoparticles immobilization: Soak the glass slide overnight in the gold nanometer solution modified with mercaptoethylamine, wash and blow dry with nitrogen.

[0030] 4. Synthesis of oligonucleotide probes: use the above-mentioned glass slides to synthesize nucleic acid sequences in an anaerobic and water-free glove box, and use the molecular stamp method to synthesize various sequences to make a chip, in which the last base is f...

Embodiment 2

[0031] Example 2, the immobilization of synthesized oligonucleotide probes and the production of non-labeled gene chips

[0032] 1. Cleaning of glass slides: Soak the slides in lotion overnight, rinse, then soak in alkali ethanol solution for two hours, rinse with double distilled water, and blow dry with nitrogen for later use.

[0033] 2. Glass slide modification: Take clean slides, soak them in the acetone solution of triethoxyaminosilane for 5 minutes, wash them, bake them at 100 degrees for 40 minutes, soak them in glutaraldehyde solution for 2 hours, and wash them with nitrogen. blow dry.

[0034] 3. Nano-gold immobilization: Soak the glass slide overnight with mercaptoethylamine-modified nano-gold solution, wash and blow dry with nitrogen.

[0035] 4. Synthesis of oligonucleotide probes: synthesized by conventional methods, one end of the oligonucleotide probe was modified with amino group, and the other end was modified with fluorescein.

[0036] 5. Chip fabrication:...

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 solid-phase nucleic acid detection probe and its preparation method are an oligonucleotide probe fixed on a solid substrate and a microarray chip made by this method, and a non-labeled oligonucleotide for detecting nucleic acid sequence information An acid probe, the probe is fixed with a fluorescence quenching material 3 through an arm molecule 2 on a solid substrate 1, and a stem composed of a fluorescent group 5 and an oligonucleotide probe molecule is prepared on the surface of the fluorescence quenching material 3. An oligonucleotide probe composed of a rod portion 6 and a ring portion 7 of the oligonucleotide probe, one end of the oligonucleotide probe is fixed on the surface of the fluorescence quenching material 3, and the other end of the oligonucleotide probe The nearby bases are marked with a fluorescent group 5, and the sequences near both ends of the oligonucleotide probe have 3 to 15 bases as complementary sequences, which enable the sequences near the two ends of the oligonucleotide probe to form In hybridization, the base sequence in the middle part of the oligonucleotide probe is the complementary sequence of the detected nucleic acid sequence.

Description

1. Technical field [0001] The invention relates to an oligonucleotide probe fixed on a solid substrate and a microarray chip made by the method, which is a non-marked oligonucleotide probe for detecting nucleic acid sequence information. 2. Background technology [0002] With the deepening of genome research, it will become possible to understand the differences in life, the rules of disease occurrence and development, and the interaction between drugs and living organisms at the genetic level. The high-throughput detection and analysis technology of nucleic acid sequence information will become one of the core technologies in the field of life sciences such as medicine. People need to develop high-throughput, accurate, and low-cost genetic information detection methods. Recently, gene chip technology has attracted more and more attention. However, there are still some problems in gene chip technology, which affect its application in biology and clinical medicine. 1) The ...

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): C12Q1/68G01N21/64
Inventor 刘全俊陆祖宏肖鹏峰
Owner SOUTHEAST UNIV
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