Fluorescence in situ hybridization probe as well as preparation method and application thereof

A fluorescence in situ hybridization and probe technology, used in biochemical equipment and methods, DNA preparation, DNA/RNA fragments, etc., can solve the problem of uncertain number of probe fluorescence, low probe specificity, and lack of small fragmentation and other problems to achieve the effect 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

Method used

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  • Fluorescence in situ hybridization probe as well as preparation method and application thereof
  • Fluorescence in situ hybridization probe as well as preparation method and application thereof
  • Fluorescence in situ hybridization probe as well as preparation method and application thereof

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preparation example Construction

[0029] The preparation method of the fluorescent in situ hybridization probe of one embodiment, comprises the steps:

[0030] Step 1: Construction of the probe library: for the non-repetitive target gene region, continuously design a series of single-stranded fragments with a length of 35nt-200nt that are completely complementary to the target gene region as candidate probes, and add The primer fragments were amplified on the chip, and the target probe library was obtained by chip synthesis.

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

[0032] In this embodiment, for different target gene regions, candidate probes are constructed at intervals of preset lengths. For example, candidate probes can be constructed at intervals of 1-10 bp, which can avoid mutual interference between probes and help improve ...

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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 ...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12N15/11
CPCC12N15/1096C12Q1/6841C12Q2563/107C12Q2543/10
Inventor 李怡
Owner 广州简册生物技术有限公司
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