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High-resolution melting method and kit for identifying animal-derived components

A technology of animal-derived components and high-resolution melting, which is applied in the field of molecular biology identification to achieve the effects of improved sensitivity, accurate detection results, and simple operation

Active Publication Date: 2019-06-11
SHAANXI UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is currently no method for high-resolution melting curve analysis using universal primers to achieve rapid, economical, and accurate analysis and identification of animal-derived components

Method used

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  • High-resolution melting method and kit for identifying animal-derived components
  • High-resolution melting method and kit for identifying animal-derived components
  • High-resolution melting method and kit for identifying animal-derived components

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Embodiment Construction

[0030] The present invention will be further described below in conjunction with drawings and embodiments.

[0031] (1) Sources of experimental materials

[0032] Fresh milk samples were collected from Caotan Dairy Farm, Weiyang District, Xi’an City; fresh goat milk was collected from the market in Weiyang District, Xi’an City; fresh buffalo milk and sheep milk were purchased from Taobao. Bead Separation Genomic DNA Extraction Kit. Non-target genomic DNA (donkey, horse, rabbit, chicken, duck, pigeon, fish, pig) were purchased from Sichuan Huahan Sanchuang Biotechnology Co., Ltd. The acquisition time is June 2018.

[0033] (2) Sequence alignment and target gene selection

[0034] The sequences of general primers selected for cattle, sheep, etc. are (GB / T 21104-2007):

[0035] Upstream primer: 5'-TTTGGTCCCAGCCTTCCTGTT-3';

[0036] Downstream primer: 5'-CTTAGTCAAACTTTCGTTT-3'.

[0037] According to the selected universal primers and the complete mitochondrial DNA sequences ...

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Abstract

The invention discloses a high-resolution melting method and kit for identifying animal-derived components. The method comprises the following steps: extracting DNA in a sample as a template, and carrying out PCR amplification by adopting a universal primer; further melting an amplification product by adopting a high-resolution melting instrument; and analyzing the shape of a high resolution melting curve to identify animal derived components in the sample. According to the invention, the target gene sequence is amplified by adopting the universal primer, so that not only can the animal-derived components in a sample be accurately identified, but also the problems of competition between primer pairs, high PCR reaction condition optimization difficulty and the like are overcome compared with the traditional multiplex PCR, and the detection efficiency is obviously improved. The method has the advantages of simplicity in operation, high detection speed, high detection sample flux and accurate detection result, and is suitable for adulteration detection and traceability identification of animal-derived components in dairy products and other foods.

Description

technical field [0001] The invention belongs to the technical field of molecular biology identification, and in particular relates to a PCR high-resolution melting method and a kit for identifying animal-derived components in foods such as dairy products. Background technique [0002] Dairy products are the main source of nutrition for a healthy diet for human beings. They are rich in protein, minerals, fat, vitamins, etc., and are deeply loved by consumers. With the improvement of living standards, people's demand for dairy products continues to increase. The dairy market has become a big sales market. Driven by economic interests, merchants use various means to reduce costs as much as possible, and the phenomenon of adulteration and sales of counterfeit products emerges in endlessly. The main adulteration behaviors are: in the production process of dairy products, shoddy and counterfeiting, such as mixing other animal milk or non-dairy raw materials with relatively low nu...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
Inventor 徐秦峰贺晓玲马西亚
Owner SHAANXI UNIV OF SCI & TECH
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