Oligonucleotide library for specific identification of whole short arm of ninth chromosome of cultivated rice and specific identification method

An oligonucleotide library and specific recognition technology, which is applied in the field of specific recognition of the whole short arm oligonucleotide library of the No. 9 chromosome of cultivated rice and can solve the problem of no specific recognition

Active Publication Date: 2017-08-29
YANGZHOU UNIV
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

At present, there is no painting probe that specifically recognizes an entire chromosome or arm in rice

Method used

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  • Oligonucleotide library for specific identification of whole short arm of ninth chromosome of cultivated rice and specific identification method
  • Oligonucleotide library for specific identification of whole short arm of ninth chromosome of cultivated rice and specific identification method
  • Oligonucleotide library for specific identification of whole short arm of ninth chromosome of cultivated rice and specific identification method

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Experimental program
Comparison scheme
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Embodiment Construction

[0018] 1. Using bioinformatics capture technology to screen 2258 oligonucleotides distributed in the entire short arm of Nipponbare chromosome 9:

[0019] Step 1: Screen and exclude all repetitive sequences in the entire short arm of chromosome 9 of the cultivated rice Nipponbare.

[0020]The second step: oligonucleotide screening: use Chorus software (https: / / github.com / forrestzhang / Chorus), (design parameters are: oligonucleotide length is 45nt, homology parameter is 75, dTm>10° (dTm=Tm-hairpinTm) oligonucleotides can be used for library construction (Tm: annealing temperature of oligonucleotides, hairpin Tm: hairpin temperature) to screen 2258 specific oligonucleotides distributed on the short arm of Nipponbare chromosome 9 Nucleotide, the sequence is shown in SEQ ID NO.1-2258.

[0021] 2. Synthesize an oligonucleotide library (2258 oligonucleotides) for probe labeling (completed by MYcroarray Company, Ann Arbor, MI). 3. Preparation of single-stranded oligonucleotide prob...

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Abstract

The invention belongs to the technical field of biology, and in particular relates to an oligonucleotide library for specific identification of a whole short arm of ninth chromosome of cultivated rice and specific identification method. The oligonucleotide library comprises total 2258 oligonucleotides shown in SEQIDNO.1-2258. The invention also discloses the specific identification method of the ninth chromosome of the cultivated rice, first, total 2258 oligonucleotide probes with fluorescent marks are prepared by use of the specific oligonucleotide library, fluorescent in-situ hybridization of the total 2258 oligonucleotide probes is performed, and the ninth chromosome of the cultivated rice can be identified according to fluorescence signal. The ninth chromosome of the cultivated rice can be identified by use of Paintes Probes of the whole chromosome arm, chromosome recombination, aberrations and homologous genes can be analyzed and studied, and the evolutionary relationship between chromosomes of different species origins can also be studied.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an oligonucleotide library for specifically identifying the whole short arm of No. 9 chromosome of cultivated rice and an identification method. Background technique [0002] Fluorescence in situ hybridization (FISH for short) is a molecular cytogenetics technique applied in the field of biotechnology. It is a non-radioactive DNA (RNA) molecular in situ hybridization technique developed on the basis of the original radioactive in situ hybridization technique in the 1980s. The basic principle of FISH is to label DNA or RNA probes with specially modified nucleotide molecules, and then directly hybridize the probes to the slices where the chromosomes or DNA fibers are located, and then use monoclonal antibodies coupled to fluorescein molecules to interact with the probes. The position of the DNA sequence on the chromosome or DNA fiber slice is detected by specific binding o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B40/06C12N15/11C12Q1/68
CPCC12Q1/6841C40B40/06C12Q2563/107C12Q2543/10
Inventor 于恒秀张韬程祝宽候莉莉沈懿龚志云顾铭洪
Owner YANGZHOU UNIV
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