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Plasmid control molecule for detection of transgenic soybean and building method thereof
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A technology of genetically modified soybean, standard molecule
Inactive Publication Date: 2010-07-14
JILIN ACAD OF AGRI SCI
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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
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[0032] Embodiment: the construction of plasmid standard molecule pMD-LM3M5
[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 DNApolymerase 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.
[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 genelectin 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 lectingene 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 enzymedigestion, 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...
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