Oligonucleotide probe and acquisition method thereof

Abstract

The invention relates to a method for acquisition of an oligonucleotide probe. The method comprises the following steps: downloading genomic sequence of crop species from a public database, filtering and finding out tandem repeat sequences, screening preliminarily found repetitive sequences, comparing residual tandem repeat sequence and known probe sequence, removing already used probe sequence, comparing residual screened tandem repeat sequences, carrying out probe design and synthesizing and verifying the designed probe sequence so as to obtain effective oligonucleotide sequence. According to the invention, cost is low and efficiency is high. The probe can be used for non-degenerated fluorescence in situ hybridization (ND-FISH) analysis of crop chromosome, determination of distribution of tandem repeat sequences represented by the probe on chromosome, understanding of the structure characteristics of chromosome and establishment of specific landmark of chromosome. Thus, specific chromosome or chromosome region of crops is identified, and the developed new oligonucleotide probe has specificity of chromosome or chromosome region.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an oligonucleotide probe and a method for obtaining the same. Background technique [0002] In crop distant hybrid breeding, allopolyploidy and plant chromosome evolution research, it is necessary to use FISH technology to analyze chromosome structure. In crop-related research, FISH technology is also essential in identifying crop chromosomes. For example, in the FISH analysis of Triticum plants, the repeated sequences pSc119.2, pAs1, pTa71, CCS1, (AAG)n and (AAC)n are commonly used probes. Recently, the Japanese have developed new repetitive sequence probes pTa-86, pTa-120p, pTa-126, pTa-465, pTa-535p, Ta-566p and Ta-713, etc. for structural analysis of wheat chromosomes. One of the important functions of repetitive sequences is to form the specific structure of chromosomes. Using repetitive sequences as probes to conduct FISH analysis on chromosomes, clarify the distr...

AI Technical Summary

Problems solved by technology

When detecting target molecules, the probe first recognizes the target sequence and forms a double-stranded structure, then under the digestion of Lambda exonuclease, the hairpin structure is opened, the G-quadruplex sequence is released, and the cycle repeats, and finally the trace target Molecular information is converted into a large number of G-quadruplex sequences, and then converted into readable optical, electrical and other signals through the G-quadruplex sequences; although the probe can realize the direct detection of single-stranded nucleic acid, it can also detect the inducible Indirect detection of single-stranded nucleic acid, double-stranded nucleic acid and other molecules (patent application number: 201610204305.3); but has the following disadvantages: (1) It can only detect single-stranded nucleic acid or double-stranded nucleic acid that can induce single-stranded nucleic acid at the DNA level and other molecules, which cannot reflect the distribution of repetitive sequences on chromosomes; (2) cannot be used to study the structure of chromosomes; (3) cannot be used for ND-FISH analysis of chromosomes

The disadvantages are: (1) focusing on single-copy sequences, which cannot reflect the distribution of repetitive sequences on chromosomes; (2) oligonucleotide probes are long, expensive to synthesize, and cumbersome to label.

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