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A universal SNP typing probe in Poaceae

A Poaceae, typing probe technology, applied in the field of molecular biology, can solve the problems of difficulty, limit the application of the Taqman probe method, and high cost of synthesizing probes, and achieve the effect of reducing the experimental cost.

Active Publication Date: 2018-10-19
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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Problems solved by technology

[0005] However, according to the principle of the Taqman probe method, a pair of specific probes needs to be synthesized for each SNP site to be identified. When there are many SNP sites to be detected, the overall cost of synthesizing probes is very high, generally very expensive. Difficult to use Taqman probe method for SNP typing
This factor severely limits the application of Taqman probe method in SNP detection

Method used

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  • A universal SNP typing probe in Poaceae
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  • A universal SNP typing probe in Poaceae

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Embodiment Construction

[0031] The present invention will be further described in detail through the following embodiments in conjunction with the accompanying drawings, but the following description does not constitute any limitation to the protection scope of the present invention.

[0032] The methods used in the following examples are conventional methods unless otherwise specified. The primers and probes used were synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.

[0033] 1. Select the wheat GW2-6A gene sequence segment with two genotypes, and the target site has two bases of C / T, see the sequence table SEQ ID NO: 5 (844bp) and SEQ ID NO: 6 ( 823bp). After amplifying and purifying this fragment, it was connected to the T vector, transformed, shaken to extract the plasmid, and finally obtained WS and OS5 plasmids with only one SNP site difference. The target site of the WS plasmid is the C genotype, and the OS5 plasmid is The T genotype is used as the typing object.

[0034] 2. Accordi...

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Abstract

The invention relates to a universal SNP typing probe in gramineous plants. The probe is composed of two pairs of reverse complement oligonucleotide sequences, and a fluorescence reporter group and a fluorescence quenching group are respectively positioned in reverse complement oligonucleotide chains. A present Taqman probe method is improved to design the SNP typing universal Taqman probe, so the experiment cost is reduced, and a Taqman probe SNP typing method can be widely applied in SNP detection of the gramineous plants.

Description

technical field [0001] The invention relates to the fields of molecular biology and nucleic acid detection, and specifically designs a general Taqman probe for SNP typing of Gramineae plants and its application in SNP site typing of Gramineae plants. Background technique [0002] Poaceae is the most economically valuable category of angiosperms, with more than 650 genera and more than 10,000 species, of which there are more than 200 genera and more than 1,200 species in my country. Poaceae plants are the main source of human food and livestock feed, have high ecological and ornamental value, and are also important raw materials for industry. [0003] Take wheat as an example. Wheat is the second largest grain crop in my country. In 2013, my country's wheat output reached 121.93 million tons. The steady growth of wheat production and continuous improvement of quality are of great significance to ensure national food security. Wheat varieties with high yield, good quality an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6827C12Q1/6858C12Q1/6895C12Q2600/156C12Q2600/166C12Q2531/113C12Q2561/101C12Q2525/191
Inventor 刘瑞娟刘宝龙刘迪车国冬蔡振媛
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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