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Nucleic acid quantitative detection kit for transgenic rice TT51-1

A technology for transgenic rice and nucleic acid quantification, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problem that the accurate quantitative detection method of transgenic rice TT51-1 has not been seen, and can ensure accuracy and traceability. The effect of high sensitivity, high sensitivity, and high sensitivity

Inactive Publication Date: 2013-10-23
NAT INST OF METROLOGY CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, the content of the precise quantitative detection method for transgenic rice TT51-1 has not yet been seen

Method used

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  • Nucleic acid quantitative detection kit for transgenic rice TT51-1
  • Nucleic acid quantitative detection kit for transgenic rice TT51-1
  • Nucleic acid quantitative detection kit for transgenic rice TT51-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Screening of internal standard gene for detection of transgenic rice TT51-1 and optimization of fluorescent quantitative PCR reaction system

[0062] 1. Materials and methods

[0063] 1. 100% transgenic rice TT51-1 rice seed powder (from China Institute of Metrology)

[0064] 2. Synthesize the primers of 5 kinds of rice internal standard genes and exogenous gene TT51-1 including GOS9, PLD, SPS, RBE4 and UBQ5 (the sequence information comes from the article published by Jeong S-C et al. in Volume 18 of Food Control magazine: 1434-1442 pages "Molecular analysis and quantitative detection of a transgenic rice line expressing a bifunctional fusion TPSP" and the article "Evaluation of four genes in rice for their suitability as endogenous reference standards in quantitative PCR") see Table 4.

[0065] 3. Using the extracted TT51-1 DNA as a template, use 5 rice internal standard gene primers and exogenous gene TT51-1 primers to carry out PCR reaction respectively....

Embodiment 2

[0091] Example 2 Quantitative detection of transgenic TT51-1 components in rice powder

[0092] 1. Materials and methods

[0093] 1. The sample to be tested is the standard substance of transgenic rice TT51-1 rice seed powder with a mass fraction of 2% (developed by China Institute of Metrology)

[0094] 2. Accurately weigh the sample to be tested and the quality control product (transgenic rice TT51-1 rice seed powder standard substance with a mass fraction of 1% and 5% content) with a balance

[0095] 3. Using the Plant Genome Extraction Kit Genomic DNA Purification Kit (Promega, USA) was used to extract the genomic DNA of the sample to be tested and the quality control product, and the purity and concentration of the extracted DNA were determined with a UV spectrophotometer and a Picogreen kit, respectively.

[0096] 4. The standard (containing about 10 6 、10 5 、10 4 、10 3 、10 2copy / ml of TT51-1 and RBE4 gene fragments (plasmid molecular standard substances) and sam...

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Abstract

The invention relates to a nucleic acid quantitative detection method and a detection kit for transgenic rice TT51-1. According to the method and the kit disclosed by the invention, the best endogenous reference gene RBE4 which has an optimal effect and is suitable for the detection of the transgenic rice is screened out; through optimization of a real-time fluorescent quantitative PCR (polymerase chain reaction) system, a PCR system for optimal quantitative detection of the transgenic rice TT51-1 or a product thereof is found. According to the method, the precise content of the transgenic component in the transgenic rice TT51-1 or the product thereof can be rapidly and correctly obtained.

Description

Technical field: [0001] The invention relates to a quantitative detection method and a detection kit for transgenic rice TT51-1 nucleic acid. Background technique: [0002] The rapid development of commercial planting and application of genetically modified crops has brought challenges to the safety management of genetically modified crops. Component testing of genetically modified products is an important part of the safety management and labeling of genetically modified organisms. [0003] The transgenic rice TT51-1 (BT63) is an insect-resistant rice, and the Ministry of Agriculture has issued a safety certificate for its production and application in Hubei Province in August 2009. There are the following reports about the transgenic detection method of TT51-1: [0004] The article "Characterization of the transgenic rice event TT51-1 and construction of a reference plasma" published in J Agric Food Chem by Cao Y. et al reported a qualitative detection method for transge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王晶隋志伟李亮余笑波臧超董莲华
Owner NAT INST OF METROLOGY CHINA
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