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Molecular marker primer for identifying Zhoushan sea area platyrhina sp and marking method

A labeling method and molecular labeling technology, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc., can solve the problem of germplasm mixing in the genus Umbrella ray

Inactive Publication Date: 2018-05-04
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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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 a marking method for identifying the undetermined species sp of the ray, which helps to solve the problem of mixed germplasm of the genus ray. The method of the present invention is simple to operate, easy to master, and accurate. high

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  • Molecular marker primer for identifying Zhoushan sea area platyrhina sp and marking method
  • Molecular marker primer for identifying Zhoushan sea area platyrhina sp and marking method
  • Molecular marker primer for identifying Zhoushan sea area platyrhina sp and marking method

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Embodiment

[0026] The undetermined species sp of the ray ray was selected from the Zhujiajian sea area of ​​Zhoushan to form a sample group of the unspecified sp. The DNA of the sample group was extracted, and PCR amplification was performed using the extracted DNA as a template. The amplification results were purified, sequenced, and compared. Sequencing results;

[0027] Described primer sequence is:

[0028] F1: 5'-TCAACCAACCACAAAGACATTGGCAC-3';

[0029] R2: 5'-ACTTCAGGGTGACCGAAGAATCAGAA-3';

[0030] The reaction system of described PCR amplification consists of:

[0031] Add in order to the 0.2ml centrifuge tube:

[0032]

[0033]

[0034] The PCR amplification program was pre-denaturation at 94°C for 5 min, (denaturation at 94°C for 35 sec, annealing at 50°C for 35 sec, extension at 72°C for 35 sec) × 38 cycles, extension at 72°C for 10 min, and preservation at 4°C. The PCR products were purified and sequenced, and the sequencing results of the two populations were compare...

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Abstract

The invention provides a molecular marker primer for identifying Zhoushan sea area platyrhina sp and a marking method. The sequence of the molecular marker primer for identifying the Zhoushan sea areaplatyrhina sp comprises a first primer and a second primer, wherein the first primer is 5'-GTTGGTAAATCTCGTGCCAGC-3', and the second primer is 5'-CATAGTGGGGTATCTAATCCTAGTTTG-3'. The invention also provides a marking method. The marking method comprises the following steps: (1) respectively extracting genomes according to selected platyrhina sp and platyrhina sinensis serving as samples; (2) takingDNA of the genomes in the step (1) as a template, designing PCR reaction systems and respectively performing amplification to obtain a molecular marker sequence of the Zhoushan sea area platyrhina spand the molecular marker sequence of the platyrhina sinensis; (3) purifying and sequencing an amplification result, comparing sequencing results, and identifying species of the Zhoushan sea area platyrhina sp.

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 marker method for identifying sp. Background technique [0002] The undetermined species of the ray, sp, belongs to the subclass Elasmobranchia, order ray, family ray, and genus ray, and was found in the sea area of ​​Zhujiajian, Zhoushan. Its external morphological characteristics are: body flat and four fan-shaped, body disk wider than body disk length, tail flat, thick and long, with a skin fold on each side, snout end blunt and round, snout cartilage short. The mouth is transversely split, the teeth are small and mostly arranged in stone pavement, with lip grooves and small eyes. There are water spray holes, about the same size as the eye diameter, and there are 2-3 longitudinal knots in the center of the back of the body from the back of the head to the front of the second dorsal fin, and there are also 2 pairs of knots ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 孙典荣张杰单斌斌陈治高天翔
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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