Primers, molecular markers and method for identifying germplasms of acipenser dabryanus and acipenser sinensis

A technology of molecular markers and Chinese sturgeon, applied in the field of molecular biology, can solve the problems that it is difficult to distinguish between two types of fish with small individuals, and it is not suitable for critically endangered species of sturgeon, and achieves low-cost, easy-to-master, and fast Effect

Pending Publication Date: 2020-03-27
FISHERIES INST SICHUAN ACADEMY OF AGRI SCI
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Problems solved by technology

[0004] The existing technology is only suitable for identifying the older two species of sturgeon, and it is difficult to distinguish the two smaller species of fish from the appearance
Moreover, the exi

Method used

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  • Primers, molecular markers and method for identifying germplasms of acipenser dabryanus and acipenser sinensis

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Example Embodiment

[0024] The preparation of sturgeon DNA is carried out by the CTAB method, and the specific steps are as follows:

[0025] Put 30mg of fin treaty into a 1.5mL centrifuge tube; add 500μL of tissue lysate (Tris-HCl 10mM, EDTA 100mM, SDS 0.6%, NaCl 400mM, 10MNaOH to adjust pH=7.5) into the centrifuge tube, and cut the tissue with clean scissors Crush, add 1μl 20mg / mL proteinase K to the centrifuge tube; water bath at 55°C for 3h, gently invert and mix every half an hour; add 120μL saturated NaCl to the centrifuge tube, mix, and place on ice for 5-10min; 4°C Centrifuge at 12000r / min for 10min, and transfer the supernatant (about 500μL) to another clean 1.5mL centrifuge tube; add an equal volume of isopropanol, shake gently, and leave for 5min; centrifuge at 12000r / min for 10min at 4℃, discard Supernatant: Add 1 mL of 70% pre-cooled ethanol to wash the DNA precipitate once, dry it naturally, and dissolve the DNA sample with an appropriate amount of double-distilled water solution; take...

Example Embodiment

[0038] Example

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Abstract

The invention discloses primers, molecular markers and a method for identifying germplasms of acipenser dabryanus and acipenser sinensis. The primers have nucleotide sequences as shown in SEQ ID NO. 3-4. The molecular markers are fragments obtained by amplification of the primers. The method comprises the following steps: amplifying genome DNA by utilizing the primers, sequencing an amplificationproduct, and comparing the amplification product with mitochondrial DNA sequences of the acipenser dabryanus and the acipenser sinensis so as to identify the acipenser dabryanus and the acipenser sinensis. The method provided by the invention has the characteristics of high speed, low cost and easiness in mastering.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a primer, a molecular marker and a method for identifying the germplasm of Acipenser dabryi and Acipenser sinensis. Background technique [0002] Chinese sturgeon (Acipenser sinensis) and Dabryanus sturgeon (A. dabryanus) are both first-class key protected aquatic wild animals in my country, belonging to the order Acipenseridae, Acipenseridae, and the genus Acipenser. They are large fishes distributed in the Yangtze River Basin. Chinese sturgeon is a large anadromous fish, an ancient and rare fish unique to my country, and one of the most primitive species of existing fish in the world. As far back as the Zhou Dynasty, more than 1000 BC, my country called Chinese sturgeon "king tuna". The Chinese sturgeon belongs to the bony fish Sturgeoniformes. It first appeared in the Early Triassic 230 million years ago and has continued to this day. It lives in the Yan...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 龚全陈叶雨赖见生刘亚宋明江
Owner FISHERIES INST SICHUAN ACADEMY OF AGRI SCI
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