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Microsatellite primers, kit and identification method for identifying tridacna crocea, tridacna squamosa and hybrid first filial generations of tridacna crocea and tridacna squamosa

The invention relates to a technology for identification methods of Saffron, which can be applied in the directions of biochemical equipment and methods, determination/inspection of microorganisms, DNA/RNA fragments, etc., and can solve the problem that hybrids are difficult to identify, and the morphological characteristics of Saffron and Scales can be solved. Indistinguishable and other problems, to achieve the effect of intuitive results, simple methods and strong reliability

Active Publication Date: 2019-12-10
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

Hybrid species are even more difficult to identify because of the indistinguishability between safflower and scale clams in the juvenile stage based on morphological characteristics

Method used

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  • Microsatellite primers, kit and identification method for identifying tridacna crocea, tridacna squamosa and hybrid first filial generations of tridacna crocea and tridacna squamosa

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Embodiment 1

[0021] A small amount of mantle tissue was cut from the foot silk holes of Safranin saffron, scaly scaly and their hybrids, and 80 individuals of each of the first generation of saffron scaly, scaly and their hybrids were extracted by phenol-chloroform extraction method Genomic DNA. Cut the adductor muscle sufficiently, absorb the water with clean filter paper, and put it into a 1.5mL centrifuge tube, then add 400μL of lysate and 10μL of proteinase K (10mg / mL), mix on a shaker, and digest in a water bath at 55℃ 3~5h, until the lysis solution is clear. Add equal volume of saturated phenol 200μL, chloroform / isoamyl alcohol mixture (24:1v / v) 200μL and extract three times. The DNA was precipitated with 1 mL of absolute ethanol, washed with 70% alcohol, dried at room temperature, and dissolved in 100 μL of ultrapure water. UV spectrophotometer was used to detect DNA concentration and purity, and 1% agarose gel electrophoresis was used to detect DNA integrity. Using the extracted ...

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Abstract

The invention discloses microsatellite primers, a kit and an identification method for identifying tridacna crocea, tridacna squamosa and hybrid first filial generations of the tridacna crocea and thetridacna squamosa. The microsatellite primers TC111 are represented as F: 5'-CCATTTTGGTGCGTAGTA-3' and R: 5'-CTTGAGGGCACCTTTTAG-3'. With the adoption of the method, the tridacna crocea, the tridacnasquamosa and the hybrid first filial generations of the tridacna crocea and the tridacna squamosa can be rapidly and accurately identified, the method has important actual application value, and meanwhile, the method has the advantages of being simple and low in cost, obtaining a direct, accurate and effective result, avoiding influence of the environment and the development stage and the like.

Description

Technical field [0001] The invention belongs to the shellfish molecular marking and hybrid identification technology in the aquatic product technology, and specifically relates to a microsatellite primer, a kit and an identification method for identifying the first generation of safflower clams, scaly clams and their hybrids. Background technique [0002] Clamshell is a large-scale marine bivalve mollusk. It not only has delicious meat and rich nutrition, but also the shells can be made into exquisite shell carving artwork, which has extremely high ornamental value, collection value and medicinal value. In recent years, due to excessive catching, clam resources have been severely damaged. For example, Coke’s clam is almost extinct in the South my country Sea. Phosphorus-free clams are also extremely rare, and most of the other clams are also endangered. Therefore, artificial clams have been carried out. Breeding and value-added release are imminent. [0003] Tridacna crocea (Tridac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2531/113C12Q2525/151C12Q2565/125
Inventor 马海涛喻子牛张跃环邓亚平肖述
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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