Method for obtaining oligonucleotide probe

An oligonucleotide probe and oligonucleotide technology, applied in the field of plant molecular chromosome engineering breeding, can solve problems such as high cost, complicated probe labeling technology, and probe labeling failure, so as to improve efficiency and avoid probe The effect of labeling failure and experimental cost reduction

Inactive Publication Date: 2014-04-09
SICHUAN AGRI UNIV
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Problems solved by technology

[0013] The purpose of the embodiments of the present invention is to provide a method for obtaining oligonucleotide probes, which aims to solve t

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  • Method for obtaining oligonucleotide probe

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Embodiment Construction

[0038] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0039] Such as figure 1 Shown, the method for obtaining oligonucleotide probe of the embodiment of the present invention comprises the following steps:

[0040] S101: Download the original sequences of pSc119.2, pAs1, pTa-535, pTa71, pAWRC.1 and CCS1 from the NCBI website;

[0041]S102: Use the bioinformatics software DNAMAN to analyze the downloaded sequences, find the repeated core units in these sequences, and then return to the NCBI website and use the blastn tool for comparison, and strive to find core oligonucleotides with a length of less than 60 bp Acid sequences for probe synthesis;

[0042] S103: Synthesizing the ...

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Abstract

The invention discloses a method for obtaining an oligonucleotide probe. The method for obtaining the oligonucleotide probe comprises the following steps: downloading original sequences of pSc119.2, pAs1, pTa-535, pTa71, pAWRC.1 and CCS1 from a national center of biotechnology information (NCBI) website; analyzing the downloaded sequences by using bioinformatics software DNAMAN; seeking repeated core units in these sequences; returning to the NCBI website to compare by using a blastn tool for finding out a core oligonucleotide sequence of which the length is below 60bp to carry out probe synthesis; carrying out probe synthesis on a lot of oligonucleotide sequences which are primarily screened out; gradually carrying out a fluorescence in situ hybridization (FISH) test after probe synthesis; verifying the function of the probe, and verifying whether the synthetic oligonucleotide sequence can play the FISH effects of pSc119.2, pAs1, pTa-535, pTa71, pAWRC.1 and CCS1 or not, so as to obtain the oligonucleotide probe. By adopting the method disclosed by the invention, complicated procedures of the existing probe marking technique are removed, the problem of marking failure of the probe is removed, and the cost is reduced.

Description

technical field [0001] The invention belongs to the technical field of plant molecular chromosome engineering breeding, in particular to a method for obtaining oligonucleotide probes. Background technique [0002] FISH technology is necessary in plant distant hybrid breeding, allopolyploidy and plant chromosome evolution research. FISH technology is also essential in the related research of Triticum family plants. Repeated sequences pSc119.2, pAs1, pTa-535, pTa71, pAWRC.1 and CCS1 are commonly used probes in FISH experiments of Triticum plants. Using these repeated sequences as probes can accurately identify the chromosomes of wheat and its close relatives, which is extremely useful for introducing foreign favorable genes into wheat in wheat breeding, and has important application value in wheat variety improvement. However, currently using these sequences as probes is time-consuming and labor-intensive. Therefore, it is necessary to improve these probes so that they can ...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/10
CPCC12Q1/6806C12Q2537/165
Inventor 唐宗祥符书兰
Owner SICHUAN AGRI UNIV
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