Method for identifying authenticity of mink fur products based on MGB probe

A technology of authenticity identification and mink, applied in the field of molecular biology, can solve the problems of high fluorescence background value and incomplete fluorescence quenching

Pending Publication Date: 2019-08-09
NORTHEAST FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fluorescence background value of the conventional TaqMan probe method is relatively high, and the fluorescence quenching is not complete

Method used

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  • Method for identifying authenticity of mink fur products based on MGB probe
  • Method for identifying authenticity of mink fur products based on MGB probe
  • Method for identifying authenticity of mink fur products based on MGB probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Detection of Suspected Mink Fur Products

[0034] 1. DNA extraction system

[0035] Take 10 mink guard hairs (average 1-2cm) and place them in a 2ml centrifuge tube, cut them into pieces, add 280μl TNE, 30μl SDS, 20μl DTT, 20μl proteinase K, add 150μl Buffer C-L (AXYGEN Animal Genome Extraction Kit), and immediately vortex Vortex for 1 min to mix well. After brief centrifugation, place the centrifuge tube in a 56°C water bath for overnight digestion. The centrifugation step was performed according to the instructions of the AXYGEN Animal Genome Extraction Kit. Add 350 μl of protein removal solution, vortex for 30 seconds to mix evenly, and centrifuge at 12000 r / min for 10 minutes. Place the DNA preparation tube in a 2ml centrifuge tube, transfer the centrifuged supernatant to the preparation tube, and centrifuge at 12000r / min for 1min. Discard the filtrate, add 500 μl washing solution, and centrifuge at 12000 r / min for 1 min. Discard the filtrate, add 700 μl desalte...

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Abstract

The invention discloses a detection method for identifying the species specificity of minks based on an MGB probe through a fluorescent PCR method. A mink upstream primer: NvF, a mink downstream primer: NvR, the mink MGB probe: NvP (the 5' end is labeled as FAM) are involved. The detection method comprises the steps of extracting DNA of sample fur; conducting fluorescent quantitative PCR amplification on the sample DNA; determining a result according to a Ct value and an amplification curve. Through a specificity experiment, a sensitivity experiment, a repeatability experiment and a blind testexperiment, the result shows that the method has high specificity and sensitivity, can be applied to quickly and accurately detecting the authenticity of mink fur products, and has wide application prospects and high practical value.

Description

technical field [0001] The invention relates to a method for identifying the authenticity of mink fur products by using a real-time fluorescent quantitative PCR method of MGB probes, belonging to the field of molecular biology. The detection method of the invention is fast and accurate. Background technique [0002] The weather in northern my country is cold, and mink fur is warm and beautiful, elegant and luxurious, and the price is also very expensive. The mink fur industry is lucrative and the processing technology is complex. It often happens that other furs are used as mink fur. However, traditional fur product inspection methods such as visual observation, scanning electron microscopy, and DNA sequencing have been unable to meet the needs of mink due to their respective limitations. The need for accurate identification of leather products. [0003] Real-time fluorescent quantitative PCR (Quantitative Real-time PCR, qPCR) is to add fluorescent groups to the convention...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 白素英徐琳雅张伟刘微张宇
Owner NORTHEAST FORESTRY UNIVERSITY
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