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Oligonucleotide probe developed by using specific sequence of elytrigia elongata

A technology of E. elongatum and oligonucleotides, applied in the field of cell biology, can solve problems such as the limitation of chromosome karyotype, and achieve the effects of abundant positions, stable hybridization signals, and accurate identification

Pending Publication Date: 2021-11-30
YANGZHOU UNIV
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Problems solved by technology

However, these probes are not derived from the specific sequences of E. elongatum, and the karyotypes analyzed are relatively limited.

Method used

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  • Oligonucleotide probe developed by using specific sequence of elytrigia elongata
  • Oligonucleotide probe developed by using specific sequence of elytrigia elongata
  • Oligonucleotide probe developed by using specific sequence of elytrigia elongata

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Embodiment 1

[0028] In order to verify the feasibility of the oligonucleotide probe Oligo-pTh1 and clarify its fluorescence signal intensity, the combination of Oligo-pTh1 and Oligo-pTh2 was used to conduct fluorescence in situ hybridization experiments in the Chinese spring-Thinopyrum elongatum addition line. The specific operations are as follows:

[0029] (1) Send the sequences of Oligo-pTh1 and Oligo-pTh2 to the company for synthesis, and modify the 5' ends of the sequences with TAMRA and FAM respectively;

[0030] (2) The synthesized probe is diluted according to the concentration requirements, and the hybridization solution is prepared according to the system in Table 1. The prepared hybridization solution is denatured in a metal bath at 103°C for 13 minutes. In cold alcohol, place it in a refrigerator above -20°C;

[0031] (3) Put the prepared chromosome sheet of the Chinese spring-Echinopsis elongatum addition line in 70% alcohol solution of 0.15mol / L NaOH for denaturation for 5min...

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Abstract

The invention relates to an oligonucleotide probe developed by using a specific sequence of elytrigia elongata, which is mainly characterized in that highly repeated sequences in elytrigia elongata are obtained by means of genome sequencing and are designed into an oligonucleotide probe Oligo-pTh1, and the oligonucleotide probe Oligo-pTh1 can be used in combination with other probes in an experiment, can enrich a chromosome FISH karyotype map, and is used for identifying structural variation of chromosomes in distant hybrid germplasm. A wheat distant hybridization material is often used as an intermediate germplasm to be applied to wheat breeding, an Oligo-pTh1 probe can assist in identifying the structural variation of chromosomes in a wheat background, the identification accuracy of exogenous chromosomes can be remarkably improved, and high-quality new wheat germplasm can be conveniently cultivated.

Description

technical field [0001] The invention relates to an oligonucleotide probe developed by using the specific sequence of Echinopsis elongatum, belonging to the field of cell biology. [0002] technical background [0003] Wheat is one of the three major food crops, providing energy and protein needed for more than 20% of the world's population. However, with the increase of the world's total population, the shrinkage of arable land, and global climate change, the wheat planting area has decreased year by year, and the parental use in breeding is single, and there is a lack of breakthrough varieties. Facing the current situation of wheat breeding, it is very important to enrich and expand the genetic basis of wheat. Practice has proved that wild relatives of wheat carry many excellent agronomic traits, and introducing these excellent genes into common wheat can broaden the genetic basis of wheat, which is one of the commonly used methods in the research of wheat genetics and bree...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6841G16B30/00
CPCC12Q1/6806C12Q1/6841G16B30/00C12Q2525/191C12Q2531/113C12Q2563/107
Inventor 戴毅石俊涛陈建民
Owner YANGZHOU UNIV
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