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A sheep meat and goat meat technology is applied in the field of molecular identification of goat meat and sheep meat, which can solve the problems of cumbersome experiments, unintuitiveness of goat meat and sheep meat, etc. Effect
Active Publication Date: 2022-03-08
INST OF AGRO FOOD SCI & TECH SHANDONG ACAD OF AGRI SCI +1
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Problems solved by technology
[0003] Aiming at the current problem of using COI to distinguish goat meat from sheep meat is not intuitive and the test is cumbersome, the present invention provides a primer pair for distinguishing goat meat from sheep meat, which can specifically distinguish goat meat from sheep meat by amplifying the mitochondrial COI gene
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Embodiment 1
[0023] Example 1 Primer pair synthesis
[0024] According to the goat and sheep mitochondrial DNA COI gene sequences published on GenBank, through BLAST comparison analysis, the Primer Premier 5.0 software was used to screen out specific primer pairs for sheep for each species-specific base site. The sequence is as shown in SEQ ID No: 1 and SEQ ID No: 2, commercially synthesized. Dilute the obtained primers to a concentration of 10 μmol / L, then aliquot them, and store them at -20°C for later use;
[0025] SEQ ID No: 1 5'-cttgtttgtatgatctgtac-3';
[0026] SEQ ID No: 2 5'-aatatgtggtgggctcatac-3'.
[0027] Embodiment 1 Differentiation of goat meat and sheep meat
[0028] (1) Use the animal genomic DNA extraction kit (Shanghai Sangong B518251) to extract the genomic DNA of the goat and sheep meat. The specific method is carried out according to the manufacturer's instructions to obtain DNA samples, and the concentration is diluted to 100 μg / mL for later use;
[0029] (2) Using...
Embodiment 2
[0034] Example 2 Specificity Verification
[0035] 2.1 Single sample
[0036] In order to detect the specificity of the primer pair, samples from goats, sheep, chickens, ducks and pigs were taken for detection. Determination method is as described in embodiment 1, and gel electrophoresis result is as follows figure 2 As shown, 1 is the DNA Marker (DL2000), 2 is the band of the amplification product of goat meat, 3 is the band of the amplification product of sheep meat, 4 is the band of the amplification product of chicken meat, and 5 is the band of the amplification product of duck meat , 6 is the band of pork amplified product. Depend on figure 2 It can be seen that only sheep meat has a bright band, and the bands of other samples are different from it. The primer can accurately and specifically distinguish sheep meat.
[0037] 2.2 Mixed samples
[0038] In order to detect the specificity of the primers, after goat meat, sheep meat, duck meat, chicken, pork were mixed...
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Abstract
The invention provides a method for distinguishing goat and sheep meat: (1) extracting the genomic DNA of the sample to be tested; (2) using the sequence shown in SEQ ID No: 1 as the upstream primer, and the sequence shown in SEQ ID No: 2 as The downstream primers were used to amplify the DNA in step (1) by PCR, and the annealing temperature was 58°C to obtain the amplified products of different samples; (3) the amplified products of different samples were subjected to agarose gel electrophoresis; (4) the results Judgment: A sample with only one amplified band is sheep meat, goat meat or other samples with multiple amplified bands, or no band. The primer pair adopted in the invention uses the sheep mitochondrial COI gene as a template, and has good specificity and high specificity. The method of using this primer pair to distinguish goat meat from sheep meat only uses a pair of primers, and can also be distinguished through the results of gel electrophoresis, without sequencing, not affected by other components, low detection cost, simple steps, and short time. Suitable for testing a large number of test samples.
Description
technical field [0001] The invention belongs to the field of molecular biology detection, and in particular relates to a molecular identification method for goat meat and sheep meat. Background technique [0002] DNA barcoding (DNA barcode) technology is a new molecular diagnostic technology for species identification that is easy to unify, easy to master, and standardized in operation. It has a good application prospect in the detection of consumer products. The study found that the mitochondrial COI gene can accurately identify and distinguish more than 13,000 species in the animal kingdom, including fish and birds. In meat products, some researchers have also used DNA barcoding technology to explore the non-directional screening technology in the adulteration of meat products, and believe that the electrophoretic bands can correctly identify a variety of animal-derived components through gene sequencing. For example, the patent application CN 105755149 A discloses the us...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2565/125
Inventor 王维婷柳尧波王守经杜鹏飞胡鹏孙苏军汝医王利斌
Owner INST OF AGRO FOOD SCI & TECH SHANDONG ACAD OF AGRI SCI
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