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Specific LAMP (Loop-mediated Isothermal Amplification) primer group, detection kit and application thereof for cows

A detection kit and specific technology, which are applied in the research field of identification of bovine-derived components detection technology, can solve the problems of large subjective influence, error, poor accuracy, etc., and achieve the effects of strong specificity, fewer samples and high sensitivity

Inactive Publication Date: 2018-05-18
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sensory detection method mainly uses visual, taste and other inspection methods to inspect food, which has the advantages of saving time, effort, and high efficiency, but there are certain errors, poor accuracy, and is greatly affected by the subjective, so it needs to be combined with other detection technologies

Method used

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  • Specific LAMP (Loop-mediated Isothermal Amplification) primer group, detection kit and application thereof for cows
  • Specific LAMP (Loop-mediated Isothermal Amplification) primer group, detection kit and application thereof for cows
  • Specific LAMP (Loop-mediated Isothermal Amplification) primer group, detection kit and application thereof for cows

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Test materials and methods

[0047] 1.1 Test material

[0048] Unless otherwise specified, all reagents were of analytical grade or biochemical reagents. Experimental water: It should meet the specifications of the first-class water in GB / T6682. DNA extraction kit: TaKaRa. Bst DNA polymerase: 8U / μl. 2x reaction buffer: 40mM Tris-HCl (pH 8.8), 20mM potassium chloride, 16mM magnesium sulfate, 1.6M betaine, dNTPs 2.8mM each. Chromogenic solution: Calcein Visual dye: Loopamp.

[0049] 1.2 Primer design

[0050] Using NCBI software to compare the mitochondrial genes of bovine-derived ingredients, select this gene sequence as the target sequence, and use the online software PrimerExplorer V5 (http: / / primerexplorer.jp / e / e) to design a primer set for loop-mediated isothermal amplification . The primer set consists of a pair of outer primers (F3, B3) and a pair of inner primers (FIP, BIP). The primer sequences are as follows:

[0051]

[0052] 1.3 Loop-mediated isothe...

Embodiment 2

[0063] Test materials and methods

[0064] 1.1 Test material

[0065] Unless otherwise specified, all reagents were of analytical grade or biochemical reagents. Experimental water: It should meet the specifications of the first-class water in GB / T6682. DNA extraction kit: TaKaRa. Bst DNA polymerase: 8U / μl. 2x reaction buffer: 30mM Tris-HCl (pH8.6), 24mM potassium chloride, 12mM magnesium sulfate, 1.4M betaine, dNTPs 2.6mM each. Chromogenic solution: 240 μM hydroxynaphthol blue.

[0066] 1.2 is the same as embodiment 1;

[0067] 1.3 Loop-mediated isothermal amplification reaction

[0068] The total volume of the loop-mediated isothermal amplification reaction system is 25ul, including 7.2ul of the loop-mediated isothermal amplification primer set solution, 14.8ul of the loop-mediated isothermal amplification reaction buffer, 1ul of Bst DNA polymerase, and the sample to be tested , 2ul each of positive and negative accusations, 2ul of hydroxynaphthol blue chromogenic solu...

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Abstract

The invention relates to the detection technical research field of identification of bovine-derived ingredients, and particularly relates to a specific LAMP (Loop-mediated isothermal amplification) primer group, a detection kit and application thereof for cows. The kit comprises the LAMP primer group, an indicator, DNA polymerase and reaction buffer liquid, wherein the LAMP primer group comprisesan external primer pair being prepared from F3 and B3 and an inner primer pair being prepared from FIP and BIP. By utilizing the detection kit, tissues of the cows and other animals can be effectively, quickly and highly specifically detected and differentiated.

Description

technical field [0001] The invention relates to the research field of detection technology for identifying bovine-derived components, in particular to a bovine-specific LAMP primer set, a detection kit and applications thereof. Background technique [0002] Dairy products are currently the main source of our country's main nutritional intake. Milk is rich in nutrients such as protein, fat and amino acids, and is a liquid product that is easily digested and absorbed by the human body. With its advantages of sweet and delicious, rich in nutrition, easy to digest and absorb, and suitable for all ages, milk has quickly entered thousands of households. But for most consumers, they may only know that milk is a good nutritional product, but they don’t know the harm of drinking adulterated milk to the human body, let alone how to identify adulterated milk. Therefore, some unscrupulous traders took this opportunity to produce adulterated milk. At present, although the company has do...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6844
CPCC12Q1/6844C12Q2531/119C12Q2563/107C12Q2537/1376
Inventor 陈爱亮李婷婷张娟王之莹张桂兰杨曙明
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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