Fluorescence in situ hybridization probe and its preparation method and application

Abstract

The invention discloses a fluorescence in situ hybridization probe as well as a preparation method and application thereof. The preparation method of the fluorescence in situ hybridization probe provided by the invention and the fluorescence in situ hybridization probe prepared with the preparation method are specifically designed for non-repetitive regions of a genome, and can reduce non-specificreactions and reduce the background interference; the constructed probe is a single-stranded DNA, and compared with traditional double-stranded probes such as BAC or PCR products, the probe providedby the invention can reduce the pairing of the probe with itself and increase the efficiency of hybridization; and the constructed single-stranded DNA probe is small in fragments, and can combine withtargeted sequences more quickly and reduce hybridization time.

Description

technical field [0001] The invention relates to the field of molecular biology detection, in particular to a fluorescent in situ hybridization probe and its preparation method and application. 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 sensitivity, long-term ...

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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