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Plasmid control molecule for detection of transgenic soybean and building method thereof

A technology of genetically modified soybean, standard molecule

Inactive Publication Date: 2010-07-14
JILIN ACAD OF AGRI SCI
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  • Summary
  • 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 patents and document reports on the construction of plasmid standard molecules for the detection of transgenic soybean MON89788

Method used

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  • Plasmid control molecule for detection of transgenic soybean and building method thereof
  • Plasmid control molecule for detection of transgenic soybean and building method thereof
  • Plasmid control molecule for detection of transgenic soybean and building method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] Embodiment: the construction of plasmid standard molecule pMD-LM3M5

[0033] 1. Experimental materials

[0034] Genetically modified soybean MON89788.

[0035] 2. Reagents

[0036] Restriction enzymes BamHI, EcoRI, and NcoI were purchased from Shanghai Sangon Bioengineering Technology Service Co., Ltd.;

[0037]The pMD18-T vector, T4 DNA ligase and buffer, ExTaq DNA polymerase and its buffer, dNTPs, and DL2000 Marker were purchased from Treasure Bioengineering (Dalian) Co., Ltd.;

[0038] PCR product recovery kit, plasmid extraction kit, and DH5α competent strain were purchased from Beijing Tiangen Biotechnology Co., Ltd.;

[0039] CTAB, Tris, EDTA and other biochemical reagents were purchased from Beijing Dingguo Biotechnology Co., Ltd.;

[0040] Primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0041] 3. Main instruments and equipment

[0042] C1000 PCR instrument (Bio-Rad Laboratories, Inc.)

[0043] Gel-type gel imaging...

experiment example

[0093] Experimental Example: Application of Plasmid Standard Molecule pMD-LM3M5 in Actual Detection

[0094] 1. Experimental materials

[0095] Genetically modified soybean MON89788.

[0096] Non-GMO soybean Ji Yu 84.

[0097]Other genetically modified materials: soybean GTS40-3-2, A5547-127, corn MON810, MON863, NK603, GA21, cotton MON1445, MON15985, MON88913, rapeseed MS1, RF1, Oxy235, beet H7-1.

[0098] 2. Reagents

[0099] 2×Mix TaqMan quantitative PCR kit was purchased from Jilin Biaoxin Technology Development Co., Ltd.

[0100] Taq DNA polymerase and its buffer, dNTPs, and DL2000 Marker were purchased from Treasure Bioengineering (Dalian) Co., Ltd.;

[0101] The plasmid extraction kit was purchased from Beijing Tiangen Biotechnology Co., Ltd.;

[0102] CTAB, Tris, EDTA and other biochemical reagents were purchased from Beijing Dingguo Biotechnology Co., Ltd.;

[0103] TaqMan probes and primers were synthesized by Shanghai Sangon Bioengineering Technology Service C...

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Abstract

The invention relates to a plasmid control molecule for detection of transgenic soybean and a building method thereof. The plasmid control molecule contains a control gene lectin specific sequence in soybean, a 3' end transformant specific sequence and a 5' end transformant specific sequence of transgenic soybean MON89788. Through designing a specific qualitative PCR primer, the lectin gene specific sequence, the 3' end transformant specific sequence and the 5' end transformant specific sequence of the MON89788 are obtained through amplification. Through molecular cloning techniques such as enzyme digestion, connection, transformation and the like, the three specific sequence segments are built into a plasmid molecule to obtain an artificially recombined plasmid control molecule pMD-LM3M5. According to tests, the plasmid control molecule built in the invention can fully substitute for the positive control of the transgenic soybean MON89788 and is suitable for the transformant specific qualitative and quantitative PCR analysis and detection of the MON89788 soybean.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a plasmid standard molecule for detection of genetically modified organisms and its products and a construction method thereof, in particular to a plasmid standard molecule suitable for qualitative and quantitative PCR detection of transgenic soybean MON89788 and a construction method thereof. Background technique [0002] Since 1996, the global planting area of ​​genetically modified crops has grown at an average annual rate of more than 10%, reaching 125 million hectares in 2008. Corresponding to the vigorous development of the industrialization of genetically modified crops, people in various countries are worried about their safety. In order to protect consumers' right to know and right to choose, many countries and regions have formulated a management system for the labeling of genetically modified organisms. Since 2001, my country has promulgated a series of laws and r...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/63C12N15/66
Inventor 李飞武张明李葱葱刘娜邢珍娟宋新元邵改革康岭生董立明夏蔚赵宁
Owner JILIN ACAD OF AGRI SCI
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