Preparation of kit and method for rapidly detecting carotene components in foods and beverages
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A technology for detection kits and carrots, which is used in biochemical equipment and methods, material excitation analysis, and microbial determination/inspection.
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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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Embodiment 1
[0038] Make the real-time fluorescent PCR detection kit of carrot ingredient by following formula, and reagent wherein comprises as follows:
[0039] (1) PCR amplification reaction solution A
[0040] Including 10×PCR reaction buffer, 0.2mmol / L dNTP (including dUTP), 2mmol / L magnesium sulfate, 0.4μmol / L upstream primer 5′-CGACAAGCAAGCTTTACTCCAA-3′, 0.4μmol / L downstream primer 5′-CGTCTGACACCCATGAGTCTGT-3 ', 0.4 μmol / L Taqman-MGB probe 5'-FAM-TCAAAACAGCCTTGAAAAACCCCACCA-TRAMA-3';
[0041] Wherein 10×PCR reaction buffer contains 100mmol / L tris-hydrochloric acid of pH8.8, 500mmol / L potassium chloride and 1% Triton X-100;
[0042] The mass ratio of the mixture of four deoxyribonucleic acids in the dNTP is dUTP:dATP:dGTP:dCTP=2:1:1:1.
[0043] (2) UNG enzyme: 1U / μL;
[0044] (3) Taq enzyme: 5 U / μL.
[0045] Follow the procedure below for testing:
[0046] (1) Extraction of DNA from carrot noodle samples to be tested
[0047] A. Weigh 0.1g of carrot noodles, cut into pieces, an...
Embodiment 2
[0065] Make the real-time fluorescent PCR detection kit of carrot ingredient by following formula, and reagent wherein comprises as follows:
[0066] (1) PCR amplification reaction solution A
[0067] Including 10×PCR reaction buffer, 0.2mmol / L dNTP (including dUTP), 2mmol / L magnesium sulfate, 0.4μmol / L upstream primer 5′-CGACAAGCAAGCTTTACTCCAA-3′, 0.4μmol / L downstream primer 5′-CGTCTGACACCCATGAGTCTGT-3 ', 0.4 μmol / L Taqman-MGB probe 5'-FAM-TCAAAACAGCCTTGAAAAACCCCACCA-TRAMA-3';
[0068] Wherein 10×PCR reaction buffer contains 100mmol / L tris-hydrochloric acid of pH8.8, 500mmol / L potassium chloride and 1% Triton X-100;
[0069] The mass ratio of the mixture of four deoxyribonucleic acids in the dNTP is dUTP:dATP:dGTP:dCTP=2:1:1:1.
[0070] (2) UNG enzyme: 1U / μL;
[0071] (3) Taq enzyme: 5 U / μL.
[0072] Follow the procedure below for testing:
[0073] (1) Extraction of the DNA of the tomato rice flour sample to be tested
[0074] A. Weigh 0.1g of tomato rice flour and tran...
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Abstract
The invention belongs to a technique for qualitatively detecting plant-derived components in foods and beverages, in particular to a kit and a method for detecting carotene components in the foods and the beverages by using a real-time fluorescent PCR (Polymerase Chain Reaction) technique. The kit comprises a PCR amplification reactive solution A, UNG (unguentum) enzyme B and Taq enzyme C, wherein the PCR amplification reactive solution A contains 10*PCR reaction buffers, magnesium sulfate, dNTP (deoxy-Nucleotide Triphosphate) containing dUTP (deoxy-Uridine Triphosphate), an upstream primer of 5'-CGACAAGCAAGCTTTACTCCAA-3', a downstream primer of 5'-CGTCTGACACCCATGAGTCTGT-3' and a TaqMan probe 5'-FAM-TCAAAACAGCCTTGAAAAACCCCACCA-TRAMA-3'. The method for rapidly detecting carotene components comprises the steps of extraction of DNA (Deoxyribonucleic Acid) in the foods and beverages, real-time PCR amplification of the carotene components and result judgement. The method has the advantages of rapidity, strong specificity, high sensitivity, convenient 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 carrot 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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