Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for identifying duck meat mixed in mutton

A technique for duck meat and mutton, which is applied in the field of PCR detection for identifying duck meat mixed with mutton, can solve the problems of high sample requirements, complicated operation, time-consuming and labor-intensive, etc., and achieve obvious band specificity and reduce false positives Effect

Active Publication Date: 2016-12-07
QINGDAO UNIV +1
View PDF10 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Driven by profit, illegal traders take advantage of the lack of food quality inspection methods to mix relatively cheap duck meat and other processed mutton into it for sale, and then make huge profits from it, which seriously violates the legitimate rights and interests of consumers.
The early identification methods were mostly based on the analysis of meat from the protein level, such as electrophoresis, enzyme-linked immunosorbent assay and other techniques, but due to complex operations, time-consuming and labor-consuming, and high sample requirements, they could not meet the requirements of fast and accurate meat identification. Require

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for identifying duck meat mixed in mutton
  • Method for identifying duck meat mixed in mutton
  • Method for identifying duck meat mixed in mutton

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Screening of amplification primers

[0028] Firstly, the most important primers in the PCR reaction were designed, and the detection sensitivity of the designed primers was verified and screened.

[0029] 1) Primer design

[0030] The mitochondrial cytochrome b gene with obvious differences in specificity between species was selected as the target gene. At the same time, in order to realize the purpose of identifying two species of mutton and duck at one time, and reduce the number of primers added in the experiment, two species Design universal degenerate primers for the highly conserved regions of the target genes of species, and select specific primers for the variable regions of each species. Design multiple groups and compare them to select the best one. Requirements: (1) The lengths of the two fragments amplified by each set of primers must be significantly different from each other, so that sheep and duck can be distinguished in electrophoresis detec...

Embodiment 2

[0041] 1. Sample preparation and DNA extraction

[0042] Take an appropriate amount of commercially available mutton and duck, mix it into mutton mixed with duck 1g / 100g, mix it well, cut out 6 portions randomly with clean tweezers or scissors, each portion is about 20mg, grind it with liquid nitrogen and place it in 6 pieces of 1.5 mL centrifuge tube. DNA was extracted with an animal tissue whole genome extraction kit (purchased from Beijing Quanshijin Biotechnology Co., Ltd.). Tissue digestion time is 2-3 hours. The OD of purified DNA was measured by UV spectrophotometer 260 / OD 280 All between 1.8-2.0, the concentration is 20-50ng / μL, 6 tubes of DNA are thoroughly mixed with each other to make a sample.

[0043] 2. PCR amplification

[0044]The 25 μL PCR reaction system contains 2.5 μL of 10-fold PCR buffer, 1 μmol / L of each of the four dNTPs, 4 μL of mixed primers, 1.25 U of Taq DNA polymerase, 100 ng of genomic DNA, and the rest is sterilized distilled water.

[004...

Embodiment 3

[0051] 1. Sample preparation and DNA extraction

[0052] Take an appropriate amount of commercially available mutton skewers, randomly cut 6 portions with clean tweezers or scissors, about 20 mg each, grind with liquid nitrogen and place them in 6 1.5mL centrifuge tubes. DNA was extracted with an animal tissue whole genome extraction kit (purchased from Beijing Quanshijin Biotechnology Co., Ltd.). Tissue digestion time is 2-3 hours. The OD of purified DNA was measured by UV spectrophotometer 260 / OD 280 All between 1.8-2.0, the concentration is 20-50ng / μL, 6 tubes of DNA are thoroughly mixed with each other to make a sample.

[0053] 2. PCR amplification

[0054] The 25 μL PCR reaction system contains 2.5 μL of 10-fold PCR buffer, 1 μmol / L of each of the four dNTPs, 4 μL of mixed primers, 1.25 U of Taq DNA polymerase, 100 ng of genomic DNA, and the rest is sterilized distilled water.

[0055] The PCR reaction program was 94°C for 5min; 32 cycles of 94°C for 30s, 50°C for ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of food quality and safety detection, and particularly relates to a PCR detection method for identifying duck meat mixed in mutton. A universal upstream primer F and two specific downstream primers R sheep and R duck are designed with mitochondrial cytochrome b gene of mutton and duck meat as characteristic target gene, a primer mixture is formed according to a certain concentration ratio, specific amplification fragments with different lengths can be obtained through one-time PCR, and analysis is conducted according to the size of molecular weight of strips obtained through electrophoresis detection. If specific strips appear on a sample to be detected at the positions of 367bp and 480bp, it proves that duck meat is mixed in the mutton sample; if specific strips appear on the sample to be detected at the position of 367bp and do not appear at the position of 480bp, it proves that no duck meat is mixed in the mutton sample. The method is accurate, reliable, simple and quick and can be used for screening species of meat in food and detecting and supervising meat adulteration.

Description

technical field [0001] The invention belongs to the technical field of food quality and safety detection, and relates to a PCR detection method for identifying duck meat mixed with mutton. Background technique [0002] Mutton is warm in nature, and in traditional Chinese medicine, it can nourish qi and tonify deficiency, promote blood circulation, and enhance resistance. It has always been regarded as one of the important foods for protecting against cold and nourishing in autumn and winter. In recent years, with the continuous improvement of living standards, the consumption of mutton and its products continues to rise, and mutton boiled, roasted mutton, and mutton pots are deeply loved by consumers. Driven by profit, unscrupulous traders take advantage of the lack of food quality testing methods to mix relatively cheap duck meat and other processed mutton into it for sale, and then make huge profits from it, which seriously violates the legitimate rights and interests of c...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143
Inventor 孙海新于金鑫孙丕春范忠刚张慧
Owner QINGDAO UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com