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Primers and probe of PCR detection for a transformant of insect resistant genetically modified cotton MON531
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A transgenic and transformant technology, applied in DNA/RNA fragments, recombinant DNA technology, fluorescence/phosphorescence, etc., can solve the problems of insufficient technology and low specificity, and achieve good versatility, sensitive detection methods, and broad application. The effect of value and market prospects
Inactive Publication Date: 2013-10-16
INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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[0005] In order to solve the above-mentioned deficiencies in the prior art, the present invention discloses primers and probes for PCR detection of insect-resistant transgenic cotton MON531 transformants and their application, mainly to realize real-time fluorescent quantitative PCR detection of transgenic cotton MON531, which overcomes the specificity of existing methods Disadvantages of weak and not advanced technology
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[0032] The primer was synthesized by Dalian Takara Company and diluted to 10μmol L -1 spare. The synthesized primer sequence is as follows:
[0033] MON531-qF:5'-ATAAAAGTATATGCGACAAAACGCG-3'
[0034] MON531-qR:5'-CTCACGTTTGTCCCTCCACTTC-3'
[0035] Extract the genetically modified cotton MON531 (1%, w / w) and genetically modified corn Bt176 (1%, w / w), MON810 (1%, w / w), T25 (1%, w / w), TC1507 (1%). ,w / w), genetically modified soybean MON87701 (1%, w / w), genetically modified rice Kefeng 6 (1%, w / w), genetically modified cotton MON1445 (1%, w / w) genomic DNA, respectively MON531 -qF / MON531-qR primer combination for SYBR Green I real-time fluorescent PCR amplification.
[0036] The extracted genetically modified cotton MON531 (1%, w / w) genomic DNA was sequentially diluted to 1%, 0.1%, 0.01%, and 0% using this as a template for SYBR Green I real-time fluorescent PCR amplification.
[0040] Primers and probes were synthesized by Dalian Takara Company, diluted to 10μmol L -1 spare. The synthesized primers and probe sequences are as follows:
[0044] Extract the genetically modified cotton MON531 (1%, w / w) and genetically modified corn Bt176 (1%, w / w), MON810 (1%, w / w), T25 (1%, w / w), TC1507 (1%). ,w / w), genetically modified soybean MON87701 (1%, w / w), genetically modified rice Kefeng 6 (1%, w / w), genetically modified cotton MON1445 (1%, w / w) genomic DNA, respectively MON531 -qF / MON531-qR / MON531-qP primer and probe combination for real-time fluorescent PCR amplification.
[0045] The extracted transgenic cotton MON531 (1%, w / w) genomic DNA was diluted to 1%, 0.1%, 0.01%, and 0% in sequence, and used as a template...
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Abstract
The invention provides primers and a probe of PCR detection for a transformant of a group of insect resistant genetically modified cotton MON531, and an application thereof. According to the invention, sequences of a forward primer and a reverse primer of the specific primers are MON531-qF:5'-ATAAAAGTATATGCGACAAAACGCG-3', and MON531-qR:5'-CTCACGTTTGTCCCTCCACTTC-3'; the sequence of the probe is MON531-qP:5'-FAM-ACCAATGCCACCCCACTGACCCA-TAMRA-3'. Through the utilization of the specific primers and the probe, a SYBR Green I real-time fluorescence PCR detection method for the genetically modified cotton MON 531 and a TaqMan probe real-time fluorescence PCR detection method are established. The two methods are sensitive, simple, and reliable, and have wide application value and market prospect.
Description
Technical field [0001] The invention relates to a method for molecular biology detection of transgenic plants in the field of biotechnology, in particular to primers and probes for PCR detection of insect-resistant transgenic cotton MON531 transformants and their applications. Background technique [0002] Genetically modified technology has brought huge benefits to agriculture and forestry production. First, genetically modified crops have high yields and rich nutritional value. Transgenic technology can increase crop yields and express substances needed by humans, such as β-carotene, to treat vitamin A deficiency. Secondly, genetically modified crops are resistant to herbicides, viruses or insect pests, which can reduce the use of chemical pesticides, reduce environmental pollution, and facilitate agricultural production and management. According to the calculation of the Environmental Impact Coefficient (EIQ), the cumulative reduction in the use of pesticides from 1996 to 20...
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IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 杨淑珂王长强路兴波孙红炜崔金杰
Owner INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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