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Molecular marker for identifying beef components and application thereof

A technology of molecular markers and beef, which is applied in the field of molecular biology, can solve problems that are not suitable for quantitative detection, and achieve the effects of improving trust, low time cost, and reducing various problems

Pending Publication Date: 2021-12-14
SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the different number of mitochondria in different cells, the mitochondrial DNA copy number will also vary in different tissues, so it is not suitable for quantitative detection

Method used

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  • Molecular marker for identifying beef components and application thereof
  • Molecular marker for identifying beef components and application thereof
  • Molecular marker for identifying beef components and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0040] Example 1DNA Extraction and Amplification Experiment

[0041] 1. Experimental materials

[0042] 1.1 Animal material

[0043] Materials for 12 different animals are: Bovine (Bovine), Pig (Sus scrofa), Sheep (Caprinae), Donkey (Equus asinus), Chicken (Gallus domesticus), Duck (Anatinae), Dog (Canis lupus familiaris), Rat (Muroidea), Fish (Piscium), Shrimp (Fenneropenaeus chinensis), Scallop (Patinopectenyessoensis), Goose (Anser cygnoides orientalis).

[0044] 1.2 Enzymes and Reagents

[0045] Hieff qPCR SYBR Green Master Mix (real-time fluorescent quantitative PCR amplification premix solution) was purchased from Shanghai Yisheng Biotechnology Co., Ltd., the catalog number is 11201ES03.

[0046] Chloroform, isoamyl alcohol, ethanol, and NaCl were of domestic analytical grade and were purchased from Sinopharm Chemical Reagent Co., Ltd. Tris, EDTA, CTAB, and SDS were purchased from Sigma-Aldrich.

[0047] The primers required for this experiment were synthesized by Wu...

Embodiment 2

[0081] Example 2 Primer Screening for Specific Amplification of Bovine Components

[0082] 1. Experimental materials

[0083] 1.1 Bovine Genomic DNA

[0084] 1.2 Enzymes and Reagents

[0085] Hieff qPCR SYBR Green Master Mix (real-time fluorescent quantitative PCR amplification premix solution) was purchased from Shanghai Yisheng Biotechnology Co., Ltd., the catalog number is 11201ES03.

[0086] Chloroform, isoamyl alcohol, ethanol, and NaCl were of domestic analytical grade and were purchased from Sinopharm Chemical Reagent Co., Ltd. Tris, EDTA, CTAB, and SDS were purchased from Sigma-Aldrich. PCR primers were synthesized by Shanghai Bioengineering Co., Ltd.

[0087] 1.3 Experimental Instruments

[0088] Real-time fluorescence quantitative PCR instrument: CFX Connect (BIO-RAD);

[0089] Ultra-micro-volume ultraviolet spectrophotometer: NanoPhotometer-N80 (Implen);

[0090] 2. Experimental method

[0091] 2.1 Design of primers

[0092] Primers were designed with the bo...

Embodiment 3

[0105] Example 3 Analysis of Species Characteristics of Primers CHR11F326-CHR11R41 for Specific Amplification of Bovine Components

[0106] 1. Experimental materials

[0107] The materials of 18 different animals and plants are: cattle (Bovine), pigs (Sus scrofa), sheep (Caprinae), donkeys (Equus asinus), chickens (Gallus domesticus), ducks (Anatinae), dogs (Canis lupus familiaris), Rat (Muroidea), fish (Piscium), shrimp (Fenneropenaeus chinensis), scallop (Patinopectenyessoensis), goose (Anser cygnoides orientalis), soybean (Glycine max), tomato (Lycopersicon esculentum), green pepper (Capsicum annuum), carrot (Daucus carota ), rice (Oryza sativa), radish (Raphanus sativus).

[0108] 2. Experimental method

[0109] 2.1. Extraction of plant genomic DNA

[0110] Plant genomic DNA was extracted using QIANGEN’s small DNA extraction kit DNeasy Plant Mini Kit (Cat. No. 69104), and the specific operations were as follows:

[0111] 1. Crush the sample and take a sample of 100mg. ...

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PUM

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Abstract

The invention discloses a molecular marker for identifying beef components and application thereof, and a beef component detection method with strong specificity and high sensitivity is established by using a cattle single copy gene in a cell nucleus as a molecular marker through a real-time fluorescent quantitative PCR technology based on SYBR Green I dye. According to the method established by the invention, cattle can be accurately distinguished from other 12 animals and 6 plants, 9 meat products on the market are actually detected, and whether the cattle-derived components are contained or not can be accurately judged. The bovine specificity real-time fluorescent quantitative PCR detection method established by the invention can provide technical reference for qualitative and quantitative detection of bovine-derived components in mixed meat products.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a molecular marker for identifying beef components in food and its application. Background technique [0002] Cattle belong to the kingdom Animalia, the phylum Chordata, the class Mammalia, the order Artiodactyla, the family Bovidae, the subfamily Bovinae, and the genus Bos. Beef is a high-quality meat food. It contains many types of essential amino acids, high content, and the amino acid composition is close to that of the human body. It has the effects of strengthening muscles and bones, nourishing the spleen and stomach, etc. With the improvement of people's recognition of the nutritional value of beef and the influence of Western food culture, the beef consumption market continues to expand; at the same time, the African swine fever in recent years has made the meat consumption structure of Chinese residents the second largest in the world after pork and poultry. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6888C12Q1/6851C12Q2531/113C12Q2563/107C12Q2527/107
Inventor 张丽杨瑶斯能武吴智华覃瑞王海英刘虹
Owner SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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