Multiplex fluorescence PCR method and kit for identifying animal-derived components

An animal-derived component, multiple fluorescence technology, applied in the field of molecular biology identification, can solve the problems of inability to obtain amplification products, unidentified identification, long target sequences, etc., to improve specificity and sensitivity, and improve detection efficiency. , the effect of preventing false negatives

Active Publication Date: 2019-06-11
SHAANXI UNIV OF SCI & TECH
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Problems solved by technology

[0005] Dairy products and other processed products, the DNA in the raw materials will be degraded into many small fragments under the influence of the processing process, which will affect the amplification primers The design puts forward very high requirements. If the target sequence amplified by the primer is too long, it is easy to fail to obtain the amplified product. If

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  • Multiplex fluorescence PCR method and kit for identifying animal-derived components
  • Multiplex fluorescence PCR method and kit for identifying animal-derived components
  • Multiplex fluorescence PCR method and kit for identifying animal-derived components

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Embodiment Construction

[0040] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0041] (1) Sources of experimental materials

[0042] 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 (duck, dog, rabbit, yak, chicken) were purchased from Sichuan Huahan Sanchuang Biotechnology Co., Ltd. Different dairy products (liquid milk, whole milk powder, skimmed milk powder and formula milk products, etc.) were purchased from Xi'an and online supermarkets. The acquisition time is June 2018.

[0043] (2) Establishment of a quadruple fluorescent PCR detection method for goat, sheep, cattle and buffalo animal-derived components in dairy products

[0044] The key to multiplex PCR amplification lies in ...

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Abstract

The invention discloses a multiple fluorescent PCR method and kit for identifying animal-derived components. The method comprises the following steps: extracting DNA in a sample as a template, carrying out multiplex PCR amplification by utilizing a plurality of pairs of designed species-specific primers, carrying out high-resolution melting curve analysis on a multiplex PCR amplification product,and judging animal-derived components contained in the sample according to a position of a melting peak of the amplified product. The method is easy to operate, short in detection period, high in specificity and reliable in detection result; the problem of pollution caused by tube opening can be avoided; and large-scale sample detection is achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology identification, and specifically relates to a method and a kit for identifying different animal-derived components such as goats, sheep, cattle, and buffaloes in raw milk and dairy products by using multiplex PCR and high-resolution melting curve analysis. Background technique [0002] With the continuous improvement of living standards, the demand for raw milk and dairy products has gradually shifted from quantity to quality, and people are more inclined to more nutritious dairy products, such as goat milk and buffalo milk with higher nutritional value. However, with the continuous increase of consumer demand, in order to prevent companies from using relatively cheap raw materials to pretend to be these dairy products with higher nutritional value for sale, countries around the world have established food labeling systems, requiring food to indicate raw material ingredients to combat adulteration....

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCY02A40/70
Inventor 徐秦峰澹台玮马西亚
Owner SHAANXI UNIV OF SCI & TECH
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