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Kit and multiplex PCR detection method for simultaneous detection of rice wide affinity gene s5 and erect panicle gene dep1

A DEP1-R and simultaneous detection technology, applied in the field of agricultural biology, can solve the problems of screening and identification of incompatible genes and erect panicle genes, and achieve the effects of simple operation, improved efficiency, and reduced costs

Active Publication Date: 2019-01-15
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can quickly and accurately identify erect panicle varieties and individual plants, but cannot simultaneously screen and identify wide-compatibility genes and erect panicle genes

Method used

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  • Kit and multiplex PCR detection method for simultaneous detection of rice wide affinity gene s5 and erect panicle gene dep1
  • Kit and multiplex PCR detection method for simultaneous detection of rice wide affinity gene s5 and erect panicle gene dep1
  • Kit and multiplex PCR detection method for simultaneous detection of rice wide affinity gene s5 and erect panicle gene dep1

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A kit for synchronous detection of rice wide affinity gene S5 and erect panicle gene DEP1, including: specific primers for wide affinity gene S5 and erect panicle gene DEP1;

[0037] The specific primers for broad affinity gene S5 are:

[0038] S5-F: 5'-GGCAATATGATATGACCAGCAC-3',

[0039] S5-R: 5'-ATGTAGCGAAGACAAAGGGAGT-3';

[0040] The specific primers for erect ear gene DEP1 are:

[0041] DEP1-F: 5'-TTTCGGTGGATCGGGTAT-3',

[0042] DEP1-R: 5'-CGTGACACCGCTAATCGT-3'.

Embodiment 2

[0044] The kit for synchronous detection of rice wide affinity gene S5 and erect panicle gene DEP1, including: 2×Es TaqMasterMix (made of Es Taq DNA Polymerase, PCR Buffer, Mg 2+ , dNTPs and PCR stabilizers and enhancers) 50mL, DNA Marker 2000, ddH 2 O 100mL, and the specific primers of broad affinity gene S5 (as shown in SEQ ID NO:1~2) and the specific primers of erect ear gene DEP1 (as shown in SEQ ID NO:3~4).

Embodiment 3

[0046] A multiplex PCR method for synchronously detecting the rice wide-compatibility gene S5 and the erect panicle gene DEP1, comprising the following steps:

[0047] 1) Material preparation

[0048] The materials to be identified are commonly used rice resources for breeding in Henan, including Yu Nongjing No. 6, Fangxin No. 1, Xinfeng No. 2, Yujing No. 5, Xindao 19, Zhengdao No. 4, Zhengdao No. 1, Zhengdao 19, Yuandao 108, Zhudan 5, Guangcan 1, Xinfeng 5, Hongguangjing 1, Zhengdao 11, Baixiangjing, Fangxin 4, Zhengjing 107, Zhengdao 5, Zhengdao 18. Zhendao 88, Yujing No. 6, Xindao 18, Jingjing No. 3, Huangjinqing, Huaidao 9, Suxiu 10, Nanjing 5055, Huaidao 6, Nanjing 42, Zhendao 99, Yan Feng 2, Yandao 11, Wuyunjing 21, Jindao 372, Nanjing 9108, Lianjing 11, Wuyungeng 23, Huaidao 8, Wuyunjing 2845, Wuyunuo 6, Wuyunuo No. 16, Zheng Han No. 9, Luodao 998, Zheng Han No. 2, Zheng Han No. 6, Upland Rice 277, Upland Rice 297, Upland Rice 502 and Xinhan No. 2, a total of 49 copie...

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Abstract

The invention discloses a kit and a multiplex PCR detection method for synchronously detecting rice broad affinity gene S5 and erect panicle gene DEP1, belonging to the field of agricultural biotechnology. According to the difference between broad affinity gene S5 and non-broad affinity, and the difference between erect ear gene DEP1 and non erect ear, the present invention designs the functional markers of the two genes. The multiplex PCR method of the gene S5 and the erect panicle gene DEP1 can detect two target genes in one PCR reaction. Compared with the single PCR method, the operation is more simple, efficient, sensitive and economical. This method is suitable for broad compatibility and erect panicle germplasm. The screening, identification and molecular aggregation breeding of resources are of great significance to improve the molecular selection efficiency of aggregation breeding and reduce the cost.

Description

technical field [0001] The invention relates to a kit for synchronous detection of rice wide affinity gene S5 and erect panicle gene DEP1, and also relates to a multiplex PCR method for synchronous detection of rice wide affinity gene S5 and erect panicle gene DEP1, belonging to the field of agricultural biotechnology. Background technique [0002] Rice is one of the important food crops in my country, and rice production accounts for about 40% of food crops. Increasing rice production is of great significance to ensuring national food security. The increase of rice yield in China mainly has two periods: the first period is the 1950s, when dwarf breeding emerged, and the second period is the widespread promotion of hybrid rice since the 1970s. Chen Wenfu, a rice expert, believes that ultra-high-yield breeding combining the use of heterosis and ideal plant type will surely be a breakthrough point for increasing rice yield in the future. [0003] There are indica subspecies ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 李俊周曹永润滕开琼赵全志杜彦修彭廷
Owner HENAN AGRICULTURAL UNIVERSITY
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