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Primer and probe for detecting garlic component in food and beverage

A technology of garlic and probes, which is applied in the direction of material stimulation analysis, microbiological determination/inspection, biochemical equipment and methods, etc., can solve the problems that the real attributes of products do not match the labels, reduce the amount of raw materials, and cannot be identified.

Inactive Publication Date: 2012-11-07
山东众合天成检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After various raw materials are processed into finished food and beverage products, it is no longer possible to identify their composition from their appearance
Producers are motivated by profit, change the composition of the product without authorization, or mix in cheap substitutes, or directly reduce the amount of raw materials, resulting in the true attributes of the product not matching the label

Method used

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  • Primer and probe for detecting garlic component in food and beverage
  • Primer and probe for detecting garlic component in food and beverage
  • Primer and probe for detecting garlic component in food and beverage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031]Make the real-time fluorescent PCR detection kit of garlic composition by following formula, and reagent wherein comprises as follows:

[0032] (1) 2×PCR Master Mix

[0033] Including 0.05u / μL Taq DNA polymerase, reaction buffer, 4mmol / L magnesium chloride, 0.4mmol / L dNTP (dATP, dCTP, dGTP, dTTP);

[0034] Wherein the reaction buffer contains 20mmol / L tris-hydrochloric acid of pH8.8, 100mmol / L potassium chloride and 2% Triton X-100;

[0035] (2) Upstream primer: 10 μmol / L, sequence: 5′-GGAAATGCTGCGAACTATGTGA-3′;

[0036] (3) Downstream primer: 10 μmol / L, the sequence is: 5′-TTGATTGGGCTGTAATGAGGC-3′;

[0037] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-TGGCAAGGAGACCTTCTGGGTTGTTAGG-3′, its 5′ end is labeled with FAM reporter fluorescent group, and its 3′ end is labeled with TAMRA quencher fluorescent group.

[0038] Follow the procedure below for testing:

[0039] (1) Extraction of DNA from samples of garlic chili sauce to be tested

[0040] A. Weigh 0.1g garlic ...

Embodiment 2

[0057] Make the real-time fluorescent PCR detection kit of garlic composition by following formula, and reagent wherein comprises as follows:

[0058] (1) 2×PCR Master Mix

[0059] Including 0.05u / μL Taq DNA polymerase Polymerase, reaction buffer, 4mmol / L magnesium chloride, 0.4mmol / L dNTP (dATP, dCTP, dGTP, dTTP);

[0060] Wherein the reaction buffer contains 20mmol / L tris-hydrochloric acid of pH8.8, 100mmol / L potassium chloride and 2% Triton X-100;

[0061] (2) Upstream primer: 10 μmol / L, sequence: 5′-GGAAATGCTGCGAACTATGTGA-3′;

[0062] (3) Downstream primer: 10 μmol / L, the sequence is: 5′-TTGATTGGGCTGTAATGAGGC-3′;

[0063] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-TGGCAAGGAGACCTTCTGGGTTGTTAGG-3′, its 5′ end is labeled with FAM reporter fluorescent group, and its 3′ end is labeled with TAMRA quencher fluorescent group.

[0064] Follow the procedure below for testing:

[0065] (1) Extraction of DNA from the bean paste sample to be tested

[0066] A. Weigh 0.1g bea...

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Abstract

The invention belongs to the field of qualitative detection technology for plant derived components in food and beverage, and specifically provides a primer / probe group for detecting garlic component in food and beverage. According to the invention, a group of specific primers and a probe are designed based on a conservative sequence of an allinase gene in garlic. The primer group and the probecan be employed in real-time fluorescent PCR technology to detect garlic component in food and beverage rapidly, sensitively and specifically. The primer group and the probe can be provided together in the form of a kit or other reagents, and applied to nucleic acid amplification reaction. The method has the advantages of simple operation and good repeatability.

Description

technical field [0001] The invention relates to a method for rapid detection of plant-derived components using nucleic acid amplification technology, in particular to a primer and probe sequence for real-time fluorescent PCR detection of garlic components. Background technique [0002] The continuous expansion of global economic integration and trade internationalization is both an opportunity and a challenge for developing China. At present, my country has become the fifth largest exporter of agricultural products in the world, and the import and export trade of agricultural products continues to grow. In order to protect their own economic interests and compete for the international market, developed countries and regions such as the United States, the European Union, and Japan have tried their best to pass various inspection items and strict or even harsh technical standards, and set up trade barriers for imported agricultural products, which has caused my country's agric...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 孙敏肖西志邓明俊龚方
Owner 山东众合天成检验有限公司
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