DNA (Deoxyribonucleic Acid) molecular identification method for Luchuan pigs and meat products thereof
A technology of DNA molecules and identification methods, which is applied in the field of DNA molecular identification of Luchuan pigs and their meat products, to achieve the effects of enriching polymorphisms, improving detection efficiency and accuracy, and requiring less templates
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Embodiment 1
[0035] Identify and distinguish Luchuan pigs and their meat products from binary cross pigs, ternary cross pigs and their meat products
[0036] Several samples of fresh meat, bacon and cut meat from Luchuan pigs, binary cross pigs, and ternary cross pigs to be identified were selected, and the DNA of each sample was extracted by the improved SDS method.
[0037] Using WM14 as primer, through ISSR-PCR reaction, the total DNA of each group of samples was amplified. ISSR-PCR reaction system is a 25μL system: template DNA 1μL, primers 2μL, 2×Taq PCR Master Mix 12.5μL, ddH 2 O9.5μL, where 2×Taq PCR Master Mix contains Taq DNA polymerase, dNTPs, MgCl 2 , Reaction buffer, PCR reaction enhancer and optimizer and stabilizer, the concentration is 2×, and does not contain dye; amplification program: 95℃ pre-denaturation for 5min, 95℃ denaturation for 30s, 36℃ annealing for 30s, 72℃ extension 2min, 30 cycles, total extension at 72℃ for 5min. Among them, the Marker is a 100bp Plus DNA ladder,...
Embodiment 2
[0044] Identify and distinguish between Luchuan pig and its meat products and Huanjiang pig fresh meat and its meat products
[0045] Several samples of fresh meat and bacon of Luchuan pig and Huanjiang pig were selected to be identified, and the DNA of each sample was extracted by the improved SDS method.
[0046] Using WM12 as the primer, ISSR-PCR amplification was the same as in Example 1.
[0047] Use 1.5% agarose gel, 1×TAE buffer as the medium electrophoresis to detect the amplification results, draw electrophoresis patterns 4 and 5 according to the electrophoresis diagram, Figure 4 The electrophoresis pattern drawing based on Picture 9 . As shown by the arrows in each figure, the Huanjiang Xiang pig samples produced a clearer band, but there is no such band in Luchuan pigs. Repeated verification experiments can prove that primer WM12 can be used to distinguish between Luchuan pigs and Huanchuan pigs. Strong evidence of Jiangxiang pig breed.
[0048] among them Figure 4 Midd...
Embodiment 3
[0053] Identify and distinguish between Luchuan pig and its meat products and Bama Xiang pig fresh meat and its meat products
[0054] Several samples of fresh meat and cut meat samples of Luchuan pig and Bama pig to be identified were selected, and the DNA of each sample was extracted by the improved SDS method.
[0055] Using WM29 as the primer, ISSR-PCR amplification was the same as in Example 1.
[0056] Use 1.5% agarose gel, 1×TAE buffer solution as the medium electrophoresis to detect the amplification results, draw electrophoresis patterns 6 and 7 according to the electrophoresis diagram, and Figure 7 The electrophoresis pattern drawing based on Picture 10 . As shown by the arrows in each figure, the Bama fragrant pig samples produced a clearer band, but there is no such band in Luchuan pigs. Repeated verification experiments can prove that primer WM29 can be used to distinguish between Luchuan pigs and Pakistani pigs. Strong evidence of Maxiang pig breed.
[0057] among th...
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