Polymerase chain reaction (PCR) detection method for trans-cry2A-genic rice line T2A-1 and application thereof

A T2A-1 and detection method technology is applied in the field of identifying transformant-specific PCR of the cry2A gene transgenic rice line T2A-1, and can solve the problems of lack of transformant-specific fragment sequences and transformant-specific detection methods, etc. High practical value, good specificity, and ineffective experimental conditions

Inactive Publication Date: 2010-11-10
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the analysis of existing patents and documents, it is found that there are no articles or patent reports on the transformant-specific fragment sequence and transformant-specific detection method of the cry2A transgenic rice line T2A-1

Method used

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  • Polymerase chain reaction (PCR) detection method for trans-cry2A-genic rice line T2A-1 and application thereof
  • Polymerase chain reaction (PCR) detection method for trans-cry2A-genic rice line T2A-1 and application thereof
  • Polymerase chain reaction (PCR) detection method for trans-cry2A-genic rice line T2A-1 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Cloning of Specific Sequence of Transgenic Rice Line T2A-1 Transgenic for cry2A

[0024] 1 Experimental materials

[0025] 1.1 Plant material

[0026] Transgenic rice: The cry2A genetically modified rice line T2A-1 (developed by Huazhong Agricultural University) is preserved in this laboratory and can be released to the public for experimental research within 20 years from the date of application.

[0027] 1.2 Enzymes and Reagents

[0028] Molecular biology reagents, such as dNTPs, Taq DNA polymerase, and DL2000Marker were purchased from Dalian Bao Biological Engineering Co., Ltd.

[0029] Other biochemical reagents are imported subpackages or domestic analytical pure. Primers were synthesized by Beijing Aoke Biotechnology Co., Ltd.

[0030] 1.3 Experimental Instruments

[0031] PCR instrument: Eppendorf Mastercycle gradient (Eppendorf co.)

[0032] Nucleic acid electrophoresis instrument: DYY-III nucleic acid electrophoresis instrument (Beijing Liuyi Instrument F...

Embodiment 2

[0057] The transformant-specific PCR method of the present invention is applied to identify the cry2A transgenic rice line T2A-1

[0058] 1 Experimental materials

[0059] 1.1 Plant material

[0060] Transgenic rice: Transgenic cry2A rice line T2A-1

[0061] 4 rice materials, purchased from the market.

[0062] 1.2 Enzymes and Reagents

[0063] Molecular biology reagents, such as dNTPs, Taq DNA polymerase, Marker, etc. were purchased from Dalian Baobio Bioengineering Co., Ltd. Other biochemical reagents are imported subpackages or domestic analytical pure. Primers were synthesized by Beijing Aoke Biotechnology Co., Ltd.

[0064] 1.3 Experimental Instruments

[0065] PCR instrument: Mastercycle gradient (Eppendorf co.)

[0066] Nucleic acid electrophoresis instrument: DYY-III nucleic acid electrophoresis instrument (Beijing Liuyi Instrument Factory)

[0067] DNA electrophoresis analysis system: Kodak EDAS 290 (Kodak co.)

[0068] Other instruments include: centrifuge, ...

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Abstract

The invention discloses a polymerase chain reaction (PCR) detection method for a trans-cry2A-genic rice line T2A-1 and application thereof, which relate to the technical field of biology. The PCR detection method for the trans-cry2A-genic rice line T2A-1 is characterized in that: a forward primer in PCR is designed according to base sequences from the first site to the 473rd site of a nucleotide sequence shown by SEQ ID NO.1, while a reverse primer is designed according to the base sequences of the 474th to 708th sites of the nucleotide sequence shown by SEQ ID NO.1. The method has the advantage of fast and accurately identifying the trans-cry2A-genic rice line T2A-1 and rice lines taking the trans-cry2A-genic rice line T2A-1 as a parent by utilizing an ordinary PCR apparatus and a reagent.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a transformant-specific PCR method for identifying the cry2A gene transgenic rice line T2A-1 and its application. Background technique [0002] As the staple food of more than half of the world's population, rice is one of the most important food crops. Over the past half century, rice breeding has achieved great success. The unit yield of rice has doubled, and even tripled in some places, which has made a huge contribution to ensuring world food security. However, the yield of rice has stagnated in the past ten years. On the one hand, there is no new breakthrough in breeding technology and the gradual narrowing of genetic diversity in cultivated varieties. On the other hand, it is also due to frequent occurrence of diseases and insect pests and droughts. Natural disasters have caused heavy losses in rice production. However, the continuous growth of the world populati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 谢家建彭于发
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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