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Primers and probes for detecting fox component in food and feed

A fox and probe technology, applied in the field of rapid detection of animal-derived ingredients, can solve problems such as unidentification, discrepancy between the real attributes of the product and the label, and reduction of the amount of raw materials

Active Publication Date: 2014-05-14
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 food and feed 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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  • Primers and probes for detecting fox component in food and feed
  • Primers and probes for detecting fox component in food and feed
  • Primers and probes for detecting fox component in food and feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Make the real-time fluorescent PCR detection kit of fox component 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 tris-hydrochloric acid, 100mmol / L potassium chloride and 2% Triton X-100 of 20mmol / LpH8.8;

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

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

[0037] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-CCCTGCACCTGGCCGGAGTC-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 the minced meat sample to be tested

[0040] A. Weigh 0.3g minced meat...

Embodiment 2

[0057] Make the real-time fluorescent PCR detection kit of fox component 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, the sequence is: 5′-TGGAGCATCAGTAGACCTTACAATTT-3′;

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

[0063] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-CCCTGCACCTGGCCGGAGTC-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) DNA extraction of the feed sample to be tested

[0066] A. Weigh 0.3g feed sample...

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Abstract

The invention belongs to technology of qualitative detection of animal-derived components in food and feed, and particularly relates to a primer / probe group for detecting fox component in food and feed. The invention selects mitochondrion cytochrome C redox enzyme I subunit gene sequence of fox, and designs a group of specific primers and probes. After using the primers and probes, the real-time fluorescent PCR (polymerase chain reaction) technology can be adopted to quickly, sensitively and specifically detect the fox component in food and feed. The primers and probes can also be provided together with other reagents in the form of a kit, and is used for nucleic acid amplification reaction. The method is simple to operate and has favorable repetitiveness.

Description

technical field [0001] The invention relates to a method for rapid detection of animal-derived components using nucleic acid amplification technology, in particular to a primer and probe sequence for real-time fluorescent PCR detection of fox components. Background technique [0002] The continuous expansion of global economic integration and trade internationalization is both an opportunity and a challenge for developing China. In recent years, my country's foreign trade has shown a sustained and steady growth trend. Statistics show that since 2009, my country has been the second largest foreign trade country in the world for three consecutive years, ranking first in export volume and second in import volume in the world. How to strengthen supervision and supervision, help and guide enterprises to improve product quality, overcome technical barriers in developed countries and regions such as the United States, the European Union, and Japan, and further expand the export of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 孙敏邓明俊肖西志高宏伟
Owner 山东众合天成检验有限公司
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