her2 gene fluorescence in situ hybridization probe and its preparation method and application

Abstract

The invention discloses a HER2 gene fluorescence in situ hybridization probe as well as its preparation method and application. The preparation method of the HER2 gene fluorescence in situ hybridization probe of the present invention and the HER2 gene fluorescence in situ hybridization probe prepared by the preparation method are specially designed for non-repeated regions of the genome, which can reduce non-specific reactions and background interference; construction The probe is single-stranded DNA. Compared with double-stranded probes such as traditional BAC or PCR products, it can reduce the pairing of the probe with itself and increase the hybridization efficiency; and the constructed single-stranded DNA probe fragments are small and can be processed more quickly Binds to targeting sequences and reduces 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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