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

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...

AI Technical Summary

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 small fragmentation, low hybridization efficiency, and long hybridization time

Moreover, when the probe is labeled with fluorescence, the fluorescence is randomly added, so the number of fluorescence of each probe is uncertain, resulting in large differences in each batch.

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