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Molecular marker primer and method for identifying Shao's Sillago and Sillago sihama

A technology of molecular markers and molecular marker sequences, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problem of germplasm mixing of genus fishes

Active Publication Date: 2017-02-22
ZHEJIANG OCEAN UNIV
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Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a molecular marker primer and method for distinguishing Shaw's carp and multi-scaled carp, which helps to solve the problem of germplasm mixing of fish of the genus Carp. The method of the present invention is simple to operate, easy to master, and accurate. high

Method used

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  • Molecular marker primer and method for identifying Shao's Sillago and Sillago sihama
  • Molecular marker primer and method for identifying Shao's Sillago and Sillago sihama
  • Molecular marker primer and method for identifying Shao's Sillago and Sillago sihama

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Embodiment 1

[0020] The Shaw's carapace sample group and the multi-scale carp sample group were composed of 3 different geographical groups, and the DNA of the 4 sample groups was extracted by selecting Shaw's carp from the coast of Xiamen, Fujian, and Changhua, Taiwan, and from the coast of Jinjiang, Fujian. Then the extracted DNA is used as a template for PCR amplification, the amplification results are purified, sequenced, and the sequenced results are compared;

[0021] The primer sequence is L5979: 5'-TTTAGTATTCGGAGCCTGAG-3'; H6241: 5'-CCTCAACACCTGATGAGGCC-3'; the reaction system of the PCR amplification consists of:

[0022] Add in order to the 0.2ml centrifuge tube:

[0023]

[0024]

[0025] The PCR amplification program was pre-denaturation at 94°C for 5 min, (denaturation at 94°C for 45 sec, annealing at 50°C for 45 sec, extension at 72°C for 45 sec) x 35 cycles, extension at 72°C for 10 min, and storage at 4°C. The PCR products were purified and sequenced, and the sequenc...

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Abstract

A molecular marker primer for identifying Shao's Sillago and Sillago sihama belongs to the technical field of biology, and sequences of the primer include L5979: 5'-TTTAGTATTCGGAGCCTGAG-3' and H6241: 5'-CCTCAACACCTGATGAGGCC-3'. The invention also provides a method for identifying Shao's Sillago and Sillago sihama by using the primer, comprising the steps of extracting DNA from Shao's Sillago and Sillago sihama, using DNAs of the two species as templates to perform PCR (polymerase chain reaction) amplification, purifying and sequencing amplification results, and comparing sequencing results to determine that intraspecific genetic difference is not greater than 1%; molecular marker sequence of Shao's Sillago is SEQ NO. 3 and molecular marker sequence of Sillago sihama is SEQ NO. 4; the method of the invention helps solve the problem of germplasm mixing of Sillago fishes, and is simple to perform, easy to master and high in accuracy.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker primer and a method for distinguishing Shaw's carpenter and multi-scale carp. Background technique [0002] Shao's catfish is a new species discovered in 2016. It belongs to the order Perciformes, the suborder Perciformes, the family Polaridae, and the genus Polarium. Currently, it is only distributed in the coastal waters of the Taiwan Strait. Its external shape is very similar to that of the squamous carp: the first dorsal fin has XI spines, the second dorsal fin has I spines and 20-22 soft rays, the anal fin has II spines and 21-22 soft rays, and has 68-73 lateral line scales ; Vertebrae number 34-37. The dorsal body of Shaw's cat is bluish-gray, the dorsal side of the head is dark, and the ventral side is white; there is a fuzzy band composed of black dots at the lower part of the lateral line on the side of the body. First and secon...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 高天翔肖家光刘璐韩志强
Owner ZHEJIANG OCEAN UNIV
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