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Method for constructing gene probe by using water-soluble conjugated polymer-as sensitized material

A technology of conjugated polymers and sensitive materials, applied in the field of bioengineering, can solve the problems of cumbersome structural design of molecular lighthouses, abnormal fluorescence enhancement reactions, configuration changes, etc., to avoid labeling and separation processes, improve molecular recognition effects, easy to combine effects

Inactive Publication Date: 2009-05-13
方圆环球光电技术盐城有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the molecular lighthouse has the advantages of high specificity, convenient operation, and time saving, there are still many defects in the molecular lighthouse technology, such as: the structural design of the molecular lighthouse is cumbersome, and the ring structure and stem structure need to be carefully designed; the choice of fluorescein Fluorescence bleaching caused by inappropriateness; the "hairpin" structure of the molecular lighthouse sometimes changes its configuration, resulting in an abnormal fluorescence enhancement reaction; the circular single-stranded structure is not as easy as the linear gene probe with the target single-stranded hybridization etc.

Method used

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  • Method for constructing gene probe by using water-soluble conjugated polymer-as sensitized material
  • Method for constructing gene probe by using water-soluble conjugated polymer-as sensitized material
  • Method for constructing gene probe by using water-soluble conjugated polymer-as sensitized material

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Embodiment Construction

[0037] Embodiment, using nano gold as fluorescence quencher, the construction method of the gene probe of immobilizing water-soluble conjugated polymer with electrostatic self-assembly technology, its flow process is as follows (see figure 1 shown):

[0038] (1) cleaning and activation of the quartz glass substrate into a negatively charged surface;

[0039] The quartz glass substrate was boiled for 10 minutes, rinsed repeatedly with deionized water three times, and dried in an oven. Then modify the surface of the substrate to activate it to be negatively charged, the steps are as follows:

[0040] ① Put the substrate into H 2 O / H 2 o 2 / NH 3 .H 3 O mixture (v / v 5:1:1) for 30 minutes;

[0041] ②Clean the substrate with deionized water or ultrasonic waves;

[0042] ③ Invade the substrate into hot H 2 o 2 / H 2 SO 4 (v / v 3:7) mixed solution for 30 minutes;

[0043] ④ Clean the substrate with deionized water or ultrasonic waves;

[0044] ⑤ Place the substrate in a dr...

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Abstract

This invention relates a method for establishing gene prodn, in which, water soluble conjguated polymer is as sensitive material, nucleic acid peptide PNA as molecular recognition element, DNA single chain as catching probe. PNA is combined to water soluble conjugated polymer. Fluorescent quencher connected to PNA molecule quenches the fluorescent of the water soluble conjugated polymer. After PNA molecule being hybridized with target single chain, the fluorescent quencher is separated from water soluble conjugated polymer, so the fluorescence of water soluble conjugated polymer is recovery, to achieve the detection of target single chain. This invention simplfies the probe establishing method, makes the hybridization of linear probe and target single chain easily. It can be used in diagnosis and bio-chemical analysis.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for constructing a gene probe using a water-soluble conjugated polymer as a sensitive material. Background technique [0002] Generally, a synthetic oligonucleotide single strand with a specific base sequence is labeled with sensitive substances for detection (such as fluorescent substances, nano gold, etc.), and the labeled strand is connected to glass, nylon membrane, nitric acid On solid support materials such as cellulose, gene probes can be constructed. When performing detection, the gene probe is hybridized with the unlabeled sample. According to the principle of complementary base pairing, the base sequence on the gene probe is hybridized and paired with the corresponding base sequence in the sample, which changes the labeling of the probe. The physical and chemical properties of sensitive substances, by detecting changes in these properties, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 黄维陈国平范曲立
Owner 方圆环球光电技术盐城有限公司
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