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Digital PCR (polymerase chain reaction) detection method for transgenic components

A technology of genetically modified components and detection methods, applied in the biological field, can solve the problems of deviation and incompatibility of experimental results, and achieve the effects of accurate detection, great flexibility, and avoidance of adverse effects

Inactive Publication Date: 2015-10-07
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current digital PCR detection methods all need to go through the sample pretreatment process, but in the actual detection, the pretreatment process often leads to deviations in the experimental results
Therefore, the existing digital PCR detection method is not suitable for direct detection of genetically modified components.

Method used

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  • Digital PCR (polymerase chain reaction) detection method for transgenic components
  • Digital PCR (polymerase chain reaction) detection method for transgenic components
  • Digital PCR (polymerase chain reaction) detection method for transgenic components

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Experimental materials and methods

[0045] 1.1 Experimental materials: The genetically modified corn samples used in this experiment: Bt176 positive samples and MIR162 positive samples, and their respective parental non-transgenic samples are all stored in the Chinese Academy of Inspection and Quarantine. The chip manufacturer is Fluidigm, the model is 48.770 Digital Array Chip, and the instrument is the BioMark system of Fluidigm Company.

[0046] 1.2 Primer design: The primers and probes used in the reaction are shown in Table 1.

[0047] Table 1

[0048] Primer, probe name

Primer, probe sequence (5'-3')

CaMV35s-F

SEQ ID No.1

CaMV35s-R

SEQ ID No.2

CaMV35s-P

SEQ ID No.3

NOS-F

SEQ ID No.4

NOS-R

SEQ ID No.5

NOS-P

SEQ ID No.6

[0049] 1.3 Genome extraction of transgenic crops

[0050] (1) The crop tissue sample is ground and thoroughly air-dried;

[0051] (2) Weigh 100 mg sample wi...

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Abstract

The invention provides a digital PCR (polymerase chain reaction) detection method for transgenic components. The digital PCR detection method includes performing digital PCR amplification reaction by the aid of primer sequences and probe sequences of the CaMV35s promoters and primer sequences and probe sequences of the NOS terminators. The digital PCR detection method has the advantages that transgenic animal and plant specimens can be accurately detected by the aid of the digital PCR detection method, and the sensitivity can reach 0.1%; steps of the digital PCR detection method are simple and are easy to operate, and accordingly the digital PCR detection method is high in flexibility during actual application and is an effective method for detecting the transgenic components.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a digital PCR detection method for genetically modified components. Background technique [0002] In 1988, the world's first glyphosate-resistant line of transgenic soybean was developed by Hinchee et al. (Hinchee, 1988). Since then, genetically modified crops have developed rapidly and have had a great impact on traditional agricultural technology. The rapid development of genetically modified crops has made more genetically modified foods flow into the commodity market, which has greatly satisfied and enriched people's material life needs; on the other hand, people have paid more and more attention to the safety of genetically modified foods. The core concern is whether foreign genes will affect the human body and the environment. Countries around the world have introduced strict management systems for genetically modified foods to prevent the possible safety concerns of genetica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 付伟朱水芳朱鹏宇王晨光许文涛张亮亮
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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