Method for identifying yak meat and cattle meat

A technology of yellow beef and yak, which is applied in the field of identifying the source of yak and yellow beef, can solve the problems of complicated operation, poor repeatability, and low sensitivity, and achieve the effect of good repeatability, high sensitivity, simple and fast operation

Active Publication Date: 2017-05-31
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above-mentioned methods have problems such as complex operation, low sensitivity, and poor repeatability,

Method used

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  • Method for identifying yak meat and cattle meat
  • Method for identifying yak meat and cattle meat
  • Method for identifying yak meat and cattle meat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Primers and probes were designed using the following 89 bp sequence in the yak mitochondrial 12S rRNA gene as the differential site; the sequence of the differential site was as follows:

[0041]

[0042] The marker primer sequences are as follows:

[0043] Upstream primer: 5'-TACCGCCATCTTCAGCAAACC-3'

[0044] Downstream primer: 5'-TTCATAGGTTACACCTTGACCTAAC-3'

[0045] The labeled probe sequence is as follows

[0046] T: 5'-GTATCATGATTGTGCTTACTTTTTTTCC-3'

[0047] 17 test samples from Gannan yak, take 200mg of each test sample, extract the whole genome DNA of the test sample, use the whole genome DNA of the sample as a template, and carry out asymmetric PCR amplification: the amplification system is as follows: upstream primer 1 μL, downstream primer 0.2 μL, probe 1 μL, sample genomic DNA 2 μL, 2×TaqMix 5 μL, ddH 2 O 1.8 μL, LC Green 1 μL. The amplification program was as follows: pre-denaturation at 95°C for 5 min; denaturation at 94°C for 30 s, annealing at 55...

Embodiment 2

[0050] From 23 samples to be tested of Pingliang Red Bull, 200 mg of each sample to be tested was taken to extract the whole genome DNA of the samples to be tested, and the rest of the method steps were the same as in Example 1.

[0051] The solubility curve of the PCR amplification product of described sample to be tested is as follows image 3 shown, according to image 3 As shown, the melting peak is in the temperature range of 65-70°C, thus it can be judged that the sample to be tested is a yellow beef sample.

Embodiment 3

[0053] 21 mixed test samples from Yushu yak and Hexi beef cattle were taken, and 200 mg of each test sample was taken to extract the whole genome DNA of the test sample. The rest of the method steps were the same as in Example 1.

[0054] The solubility curve of the PCR amplification product of described sample to be tested is as follows Figure 4 shown, according to Figure 4 As shown, there are melting peaks in the temperature range of 55-60°C and 65-70°C, so it is judged that the sample to be tested is a mixed sample of yak meat and yellow beef.

[0055] It can be seen from the above examples that the method of the present invention adopts the HRM method to operate simply and quickly, and only needs 2 hours to complete a detection. It has high sensitivity and good repeatability, and can accurately detect a large number of samples at the same time.

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PUM

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Abstract

The invention provides a method for identifying yak meat and cattle meat by adopting an HRM method. The method comprises the following steps: (1) selecting different loci of the mitochondrial 12S rRNA gene of the yak and the mitochondrial 12S rRNA gene of the cattle, and designing a marking primer and a marking probe according to the selected different loci; (2) extracting the whole genome DNA of a to-be-detected sample; (3) by taking the whole genome DNA of the to-be-detected sample obtained in the step (2) as a template, adding with the marking primer and the marking probe in the step (1), and carrying out asymmetrical PCR amplification, thus obtaining a PCR amplification product; and (4) detecting the melting temperature of the PCR amplification product obtained in the step (3) by adopting an HRM method, and drawing a melting curve, wherein if the melting peak is within 55-60 DEG C, the to-be-detected sample is detected to be the yak meat, and if the melting peak is within 65-70 DEG C, the to-be-detected sample is detected to be the cattle meat.

Description

technical field [0001] The invention relates to the field of meat quality detection, in particular to a method for identifying the origin of yak and yellow meat. Background technique [0002] Meat adulteration has become one of the important challenges faced by my country's food and meat quality and safety control. Yak meat is a pollution-free green food unique to the plateau. Because of its high nutritional value and unique flavor, the corresponding price is also quite different from other beef. Phenomenon. [0003] At present, the routine identification methods for meat-derived components mainly use PCR-based nucleic acid level detection, including DNA probe hybridization, RFLP-PCR (restriction fragment length polymorphism amplification), RAPD-PCR (random Amplified polymorphism PCR) and other methods. However, the above-mentioned methods have problems such as complex operation, low sensitivity, and poor repeatability, which are obstacles to the rapid and accurate identi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2527/107
Inventor 郭婷婷
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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