HER2 gene fluorescent in-situ hybridization probe as well as preparation method and application thereof

A fluorescence in situ hybridization and probe technology, applied in biochemical equipment and methods, DNA preparation, DNA/RNA fragments, etc., can solve the problem of uncertain number of fluorescent probes, low probe specificity, and lack of small fragmentation etc. to achieve the effects of reducing background interference, increasing hybridization efficiency, and reducing hybridization time

Active Publication Date: 2019-01-08
广州简册生物技术有限公司
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Problems solved by technology

However, there are some non-specific sequences in the BAC and PCR product probes, the probe specificity is not high, and the signal background is high
Existing probe fragments are generally too large, generally between 200-500nt in length, lack of effective methods for s

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  • HER2 gene fluorescent in-situ hybridization probe as well as preparation method and application thereof
  • HER2 gene fluorescent in-situ hybridization probe as well as preparation method and application thereof
  • HER2 gene fluorescent in-situ hybridization probe as well as preparation method and application thereof

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Example Embodiment

[0029] The preparation method of the HER2 gene fluorescent in situ hybridization probe of one embodiment includes the following steps:

[0030] Step 1: Construct the probe library: select the non-repetitive target gene region on chromosome 17 of the human genome containing the HER2 gene, and construct a 35nt-200nt single-stranded fragment that is completely complementary to the target gene region every 1-10bp as Candidate probes, through BLAST comparison, obtain the Tm value of non-specific hybridization, remove the non-specific Tm value for candidate probes at 45°C, and select non-overlapping candidate probes, and select the candidate probes at both ends of the screen. Add the amplified primer fragments to obtain the target probe library.

[0031] The target gene region in this embodiment is a non-repetitive region of the genome. By designing candidate probes for the non-repetitive region, non-specific reactions and background interference can be reduced.

[0032] For different tar...

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Abstract

The invention discloses an HER2 gene fluorescent in-situ hybridization probe as well as a preparation method and application thereof. The preparation method of the HER2 gene fluorescent in-situ hybridization probe as well as the HER2 gene fluorescent in-situ hybridization probe prepared with the preparation method are specially designed for a genome non-repeated region, so that non-specific reaction can be reduced and background interference can be reduced; the constructed probe is single-chain DNA, and can reduce pairing of the probe and the own and improve the hybridization efficiency compared with the traditional double-chain probe such as BAC or PCR products; furthermore, the constructed single-chain DNA probe has a small segment, can be combined with a targeted sequence rapidly and can reduce the hybridization time.

Description

technical field [0001] The invention relates to the field of molecular biology detection, in particular to a HER2 gene fluorescence in situ hybridization probe and a preparation method and application thereof. Background technique [0002] Fluorescence in situ hybridization (FISH) is based on known population-specific DNA sequences, uses fluorescently labeled DNA fragments as probes, and hybridizes with DNA molecules in the cell genome to detect the existence and abundance of the specific DNA. Spend. The basic principle of FISH is to label DNA (or RNA) probes with special nucleotide molecules, and then directly hybridize the probes to chromosomes or DNA fiber slices, and then use monoclonal antibodies conjugated to fluorescein molecules to interact with the probes. Qualitative, localization, and relative quantitative analysis of DNA sequences on chromosomes or DNA fiber slices by specific binding of needle molecules. FISH has the advantages of safety, rapidity, high sensit...

Claims

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Application Information

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IPC IPC(8): C12Q1/6841C12Q1/6811C12N15/11C12N15/10
CPCC12Q1/6811C12Q1/6841C12Q2563/107C12Q2531/113C12Q2563/159
Inventor 李怡
Owner 广州简册生物技术有限公司
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