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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 existing technology to identify germplasm from the number and shape of gill rakers requires killing fish and dissecting it, which is not suitable for critically endangered protected species such as sturgeon

Method used

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

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preparation example Construction

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

[0025] Put about 30 mg of fins into a 1.5 mL centrifuge tube; add 500 μL of tissue lysate (Tris-HCl 10 mM, EDTA 100 mM, SDS 0.6%, NaCl 400 mM, 10M NaOH to adjust pH = 7.5) into the centrifuge tube, cut the tissue with clean scissors Add 1μl 20mg / mL proteinase K to the centrifuge tube; bathe in 55℃ water for 3h, gently invert and mix every half hour; add 120μL saturated NaCl to the centrifuge tube, mix well, place on ice for 5-10min; 4℃ , centrifuge at 12,000r / 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 let stand for 5min; centrifuge at 12,000r / min for 10min at 4°C, discard Supernatant: Add 1mL of 70% pre-cooled ethanol to wash the DNA pellet once, and after natural drying, dissolve the DNA sample with an appropriate amount of double-distilled aqueous solut...

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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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