A strain-specific qualitative and quantitative PCR detection method for transgenic carnation moonlite

A detection method, carnation technology, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., to achieve high sensitivity and strong specificity

Inactive Publication Date: 2011-12-21
SHANGHAI ACAD OF AGRI SCI
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At present, domestic scientific research institutions have not ...

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  • A strain-specific qualitative and quantitative PCR detection method for transgenic carnation moonlite
  • A strain-specific qualitative and quantitative PCR detection method for transgenic carnation moonlite
  • A strain-specific qualitative and quantitative PCR detection method for transgenic carnation moonlite

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Embodiment Construction

[0043] The technical solution of the present invention will be further described in detail below in conjunction with specific embodiments.

[0044] The present invention takes transgenic carnation Moonlite strain specific detection research as an example,

[0045] The specific method is as follows:

[0046] 1. Extract the genomic DNA of transgenic carnation Moonlite.

[0047] Genomic DNA of transgenic Carnation Moonlite was extracted using the plant genome extraction kit "Plant DNA Mini-prep kit" (Ruifeng Biotechnology Co., Ltd., Shanghai). Utilize spectrophotometer (Thermo Company EV60) to measure DNA concentration and purity to be 10ng / μL-100ng / μL and OD260 / 280=1.7-2.0 respectively, according to carnation genome DNA size (Royal botanic Gardens, Kew.Plant c-DNA value The database (release 4.0, October 2005) determined its copy number to be 15873-158730 copy / μL.

[0048] 2. Amplification and confirmation of the adjacent sequence on the left side of the foreign gene insertio...

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Abstract

The invention discloses a qualitative and quantitative PCR (polymerase chain reaction) detection method for strain specificity of transgenic carnation Moonlite, which comprises the following steps: 1) extraction of genome DNA (deoxyribonucleic acid) of the transgenic carnation Moonlite; 2) amplification and confirmation of the left boundary adjacent sequence of the transgenic carnation Moonlite exogenous gene insertion site; 3) establishment and verification of the qualitative PCR detection method for transgenic carnation Moonlite; and 4) establishment and verification of the quantitative PCRdetection method for transgenic carnation Moonlite. The invention successfully establishes the qualitative and quantitative PCR detection method for strain specificity of transgenic carnation Moonlite, and verifies the specificity and sensitivity of the detection method, thereby providing references for import detection, supervision and environmental safety evaluation of transgenic carnation.

Description

technical field [0001] The invention belongs to the technical field of transgenic plant detection, and in particular relates to a strain-specific qualitative and quantitative PCR detection method of transgenic carnation Moonlite. Background technique [0002] Since the first transgenic tomato FLAVR SAVR was approved by the USDA for commercial production in 1994, more and more transgenic plants have been used in agricultural production. The industrialization of genetically modified plants, especially the industrialization of genetically modified crops can reduce the impact of agriculture on the environment, increase food production, and help solve the global food problem. According to statistics, during the 15 years from 1996 to 2010, the total area of ​​GM crops planted exceeded one billion hectares. The planting area of ​​genetically modified crops increased 87 times from 17,000 hectares in 1996 to 1.48 million hectares in 2010. In 2010, the number of countries planting g...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 潘爱虎李鹏贾军伟蒋玲曦朱宏白蓝王金斌唐雪明
Owner SHANGHAI ACAD OF AGRI SCI
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