Method for precisely identifying golden carp by utilizing mitochondrial NADH5 genes
A mitochondrial and genetic technology, applied in the biological field of identifying fish species, can solve problems such as difficult identification through appearance distinction
Active Publication Date: 2018-11-20
广西壮族自治区水产引育种中心
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Problems solved by technology
However, only about 10% of the individuals in this population have golden skin from the upper edge of the head to the upper edge of the tail, which forms a thick golden line continuously or intermittently, like a "golden edge" on the back of the fish, and the remaining 90% of the individuals The shape of the carp is similar to many geographical populations and artificially selected species of carp, especially at the species stage, it is difficult to identify through the shape difference. Provide accurate identification basis for germplasm identification and disputes arising from commercial breeding
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Embodiment 1
[0060] The method of using the mitochondrial NADH5 gene to accurately identify the Phnom Penh carp includes the following:
[0061] 1. Extract the DNA of Phnom Penh carp to be identified, and use the TIANGEN Genomic DNA Extraction Kit to extract, specifically including the following steps:
[0062] (1) Mince a small amount of fish fin tissue (about 10mg) into a 1.5mL centrifuge tube, add 200μL GA solution and mix well;
[0063] (2) Add 20 μL proteinase K solution and mix well, digest in a water bath at 56°C until the tissue dissolves (about 3 hours);
[0064] (3) Add 200 μL of GB buffer, shake thoroughly by inverting, and place in a water bath at 70°C for 10 minutes;
[0065] (4) Add 200 μL of absolute ethanol, shake and mix well for 15 s, transfer the solution to the adsorption column CB3, put the CB3 adsorption column into the collection tube, centrifuge at 12000 rpm for 30 s, discard the waste liquid, and put the adsorption column back into the collection tube;
[0066] (...
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The invention discloses a method for precisely identifying golden carp by utilizing mitochondrial NADH5 genes. The method comprises the following steps: (1) extracting DNA of to-be-identified golden carp; (2) performing PCR on an individual DNA template of the golden carp by utilizing primers; (3) recovering the amplification product; (4) cloning and sequencing the PCR amplification products of the primers; (5) performing comparative analysis on the sequences. According to the method for identifying the golden carp by utilizing mitochondrial NADH5 genes disclosed by the invention, the similarity between the sequence of the NADH5 genes of the golden carp and the carp sequence submitted in the invention is 99%, that is, the golden carp belongs to one of the carps; however, bases of the site247 and the site 559 in the sequence of the golden carp are respectively A and C, and are different from bases G and T at corresponding sites of other carps, so that the golden carp can be precisely identified. The golden carp is identified by using the method disclosed by the invention, defects of morphological identification are overcome, and a reference is provided for protecting the local variety of the golden carp and solving commercial dispute of aquaculture species.
Description
technical field [0001] The invention belongs to the field of biotechnology for identifying fish species, and specifically relates to an accurate method for identifying Phnom Penh carp and other carp, which is actually a method for accurately identifying Phnom Penh carp by utilizing the mitochondrial NADH5 gene. Background technique [0002] Scientific name of carp: Cyprinus carpio . It is native to Asia and later introduced to Europe, North America and other regions. It is omnivorous. The scales are large, and there are two whiskers on each side of the palate. They live alone or in small groups in calm mud-bottom ponds, lakes, and rivers overgrown with water and grass. From the perspective of nutritional value, the protein content of carp is not only high, but also of good quality. The human body's digestion and absorption rate can reach 96%, and it can supply essential amino acids, minerals, vitamin A and vitamin D; every 100g of meat contains protein. 17.6g, 4.1g of fat...
Claims
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IPC IPC(8): C12Q1/6869C12Q1/6888C12N15/70
CPCC12N15/70C12Q1/6869C12Q1/6888C12Q2531/113
Inventor 叶香尘韦玲静吕业坚文衍红张桂姣黄杰杨著山张盛滕忠作甘宝江
Owner 广西壮族自治区水产引育种中心