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Nuclear acid probe marked with quantum-dots composite sphere and preparation method and application thereof

A technology of nucleic acid probes and quantum dots, which is applied in the fields of molecular biology and cytogenetics, can solve problems such as large interference between spectra, weak fluorescence intensity, and excitation of different wavelengths, and achieve strong fluorescence stability, stable probes, The effect of the simple preparation method

Inactive Publication Date: 2009-09-09
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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Problems solved by technology

[0006] In view of this, one of the objectives of the present invention is to provide a quantum dot combined microsphere-labeled nucleic acid probe, which can overcome the weak fluorescence intensity, easy photobleaching, and different fluorescent dye-labeled nucleic acid probes existing in the existing fluorescent dye-labeled nucleic acid probes. Nucleic acid probes need to be excited at different wavelengths and have large interference between spectra, etc.

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  • Nuclear acid probe marked with quantum-dots composite sphere and preparation method and application thereof

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preparation example Construction

[0028] There are two main methods for preparing quantum dots: one is prepared in an organic phase, and the other is prepared in an aqueous phase. Quantum dots with small particle size distribution and high fluorescence quantum yield can be prepared by colloidal chemistry in the organic phase, but they are insoluble in water. When applied to nucleic acid labeling, certain bifunctional groups must be used first. Its surface is modified so that it can be coupled with nucleic acid molecules while being water-soluble. There are multiple methods for preparing water-soluble quantum dots: (1) ligand exchange method, replace the tri-n-octyl phosphine oxide / tri-n-octyl phosphine ( TOPO / TOP) ligand, this bifunctional ligand is mainly composed of two parts, one part can be fixed on the surface of quantum dots (such as -SH), and the other part is a hydrophilic group (such as -OH, -COOH) (2) Loading method, such as loading quantum dots through a polymerized silicon shell, the inner layer o...

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Abstract

The invention discloses a nuclear acid probe marked with quantum-dots composite sphere and a preparation method and application thereof. Nuclear acid probes with different sequences are marked with quantum-dots composite sphere with different emission spectrum characteristics; the quantum-dots composite sphere comprises a sphere and a quantum-dots composite contained in the sphere, the quantum-dots composite is composed of randomly selected m types of quantum-dots with different emission spectrum characteristics from n types thereof, n and m are both positive whole numbers and m is less than or equal to n; the preparation method of the nuclear acid probe comprises the following steps: preparing numerous probes with different sequences and marking the probes with X; preparing numerous quantum-dots composite sphere with different emission spectrum characteristics and marking the sphere with Y; integrating probes marked with X and quantum-dots composite sphere marked with Y through specificity of X and Y and carrying out coupling on the probes and sphere; applied to multicolor fluorescence in situ hybridization, spectral karyotype analysis and chromosome karyotype analysis, the probeof the invention can improve sensitivity, repetitiveness and specificity of detection, simplify detection device, increase detection speed and prevent false positive result from occurring.

Description

technical field [0001] The invention relates to the fields of molecular biology and cytogenetics, in particular to a quantum dot combined microsphere labeled nucleic acid probe and a preparation method and application thereof. Background technique [0002] Chromosomes of different species have their own specific morphological and structural characteristics (including chromosome length, centromere position, arm ratio, satellite size, etc.), and this morphological and structural characteristics are relatively stable. Therefore, karyotype analysis is an important content of genetic research, and it is also of great scientific significance for the study of the occurrence, development, and metastasis mechanisms of human chromosomal diseases and malignant tumors. [0003] The traditional method of chromosome karyotype analysis is mainly the chromosome G banding method, which has the advantages of simple operation and low cost, and can accurately distinguish the deletion, duplicati...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10G01N21/64
Inventor 罗阳王珏府伟灵
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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