A DNA molecular identification method for Luchuan pig and its meat products
A DNA molecule and identification method technology, which is applied in the field of DNA molecular identification of Luchuan pigs and their meat products, achieves the effects of rich polymorphism, improved detection efficiency and accuracy, and simple operation
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Embodiment 1
[0035] Identify and distinguish Luchuan pig and its meat products from binary miscellaneous pigs, triple miscellaneous pigs and their meat products
[0036] Several samples of raw meat, bacon and buckwheat from Luchuan pig, binary mixed pig, and triple mixed pig were selected for identification, and the DNA of each sample was extracted by the improved SDS method.
[0037]Using WM14 as a primer, the total DNA of each group of samples was amplified through ISSR-PCR reaction. The ISSR-PCR reaction system is 25 μL system: template DNA 1 μL, primer 2 μL, 2×Taq PCR Master Mix 12.5 μL, ddH 2 O9.5μL, in which 2×Taq PCR Master Mix contains Taq DNA polymerase, dNTPs, MgCl 2 , reaction buffer, PCR reaction enhancer, optimizer and stabilizer, the concentration is 2×, and does not contain dye; amplification program: 95°C pre-denaturation for 5min, 95°C denaturation for 30s, 36°C annealing for 30s, 72°C extension 2min, cycled 30 times, total extension at 72°C for 5min. Among them, Marker...
Embodiment 2
[0044] Identification and distinction of Luchuan pig and its meat products from fresh meat of Huanjiang Xiang pig and its meat products
[0045] Several fresh meat samples of Luchuan pig, Huanjiang Xiang pig and their bacon were selected to be identified, and the DNA of each sample was extracted by the improved SDS method.
[0046] Using WM12 as a primer, the ISSR-PCR amplification was the same as in Example 1.
[0047] Use 1.5% agarose gel and 1×TAE buffer as the medium to electrophoresis to detect the amplification result, draw the electrophoresis pattern 4 and 5 according to the electropherogram, and use Figure 4 The electrophoretic pattern drawn based on Figure 9 . As shown by the arrows in each figure, a relatively clear band was produced in Luchuan pig samples, but there was no such band in Huanjiang Xiang pigs. Repeated verification experiments can prove that the primer WM12 can be used to distinguish Luchuan pigs from Huanjiang pigs. Strong evidence for the Jiangx...
Embodiment 3
[0053] Identification and distinction of Luchuan pig and its meat products from Bamaxiang pig fresh meat and its meat products
[0054] Several samples of Luchuan pig, Bamaxiang pig fresh meat and their braised meat samples were selected, and the DNA of each sample was extracted by the improved SDS method.
[0055] Using WM29 as a primer, the ISSR-PCR amplification was the same as in Example 1.
[0056] Use 1.5% agarose gel and 1×TAE buffer as the medium to electrophoresis to detect the amplification results, draw the electrophoretic patterns 6 and 7 according to the electrophoretic graphs, and use Figure 7 The electrophoretic pattern drawn based on Figure 10 . As shown by the arrows in each figure, a relatively clear band was produced in the samples of Bamaxiang pigs, but there was no such band in Luchuan pigs. Repeated verification experiments can prove that the primer WM29 can be used to distinguish Luchuan pigs from Bama pigs. Strong evidence for the horse-scented pig...
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