Identification method of single nucleotide polymorphism of plectropomus leopardus in different groups
A technology of single nucleotide polymorphism and leopard gillfish, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of difficult and accurate identification, high adult price, etc., and achieve easy operation and identification method Easy and fast effect
Inactive Publication Date: 2013-11-13
XIAMEN UNIV
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At present, the East Star Spot is cultivated in Hainan and Guangdong, while the Red East Star Spot is the first choice for cultivating seedlings because of its high adult price. The price is significantly higher than that of the White East Spot Spot. The morphological characteristics are very similar, but it is difficult to accurately identify them at the stage of fertilized eggs and seedlings (the real body color has not yet formed), so many lawbreakers are given a shoddy opportunity to replace Hongdongxing spotted seedlings or fertilized eggs with Baidongxing spotted seedlings or fertilized eggs. Eggs are sold to farmers for profit
At present, there is no official report on the method for accurately distinguishing red and white eastern star seedlings
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[0022] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.
[0023] 1) The selected samples are the individual muscle samples of red eastern star spot and white eastern star spot, and extract them by phenol chloroform extraction method according to the conventional method (see "Molecular Cloning Experiment Guide" (third edition) translated by Huang Peitang et al., Science Press, September 2002) to extract Eastern star spot DNA;
[0024] 2) Design and synthesize a pair of selective amplification primers
[0025] COI-606H: 5'-CAGTATCTTTCTCTCTATCACTACCTGTTATA-3'
[0026] COI-606L: 5'-AATAAGTGCTGGTAGAGGATTGGGT-3'.
[0027] The mitochondrial DNA gene of Dongxingban was amplified by PCR twice, and the amplification procedure for the positive detection reaction was as follows: pre-denaturation at 95°C for 5 minutes; the first 35 cycles were denaturation at 95°C for 30s, annealing at 58°C for 30s, and annealing at 72...
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The invention relates to an identification method of single nucleotide polymorphism of plectropomus leopardus in different groups, and relates to plectropomus leopardus. The identification method comprises the following steps: 1) extracting a genome DNA (Deoxyribonucleic Acid) of Miniatus Grouper; 2) by using the genome DNA obtained in the step 1) as a template, performing PCR (Polymerase Chain Reaction) twice to amplify the COI (Cytochrome Oxidase I) gene in a mitochondria genome so as to obtain a PCR amplification product; and 3) performing electrophoretic analysis on the PCR amplification product, wherein if the PCR amplification product at 104bp is detected, the sample is red Miniatus Grouper, and if the PCR amplification product at 104bp is not detected, the sample is white Miniatus Grouper. The identification method is simple, convenient and fast, and easy to operate.
Description
technical field [0001] The invention relates to leopard echinacea, in particular to a method for identifying single nucleotide polymorphisms of different groups of leopard echinacea. Background technique [0002] Plectropomus leopardus, also known as Eastern star spot, belongs to Perciformes (Perciformes), grouper (Epinephelidae), genus Plectropomus, and its wild populations are widely distributed in the Indian Ocean-West Pacific The region, from southern Japan in the north, to Australia in the south, and to Fiji in the east, is a famous and precious seafood in the southeast coast of my country and even in the world. At present, there are two types of East Star Spot on the market, one of which is ruddy in color, commonly known as Red East Star Spot, because it is full of festive colors, it is in line with the shopping preferences of Chinese people, and the price is expensive, about 500 yuan / kg, while the other is gray and white. Commonly known as Baidong star spot, its price...
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IPC IPC(8): C12Q1/68
Inventor 蔡晓鹏王航俊丁少雄曲朦张荣墨
Owner XIAMEN UNIV