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A kind of identification primer and method suitable for giant clam oyster, common species of giant clam in the South China Sea and its hybrids

An identification method and technology of giant clam, applied in biochemical equipment and methods, measurement/inspection of microbes, DNA/RNA fragments, etc. and the effects of developmental period, intuitive results, and simple methods

Active Publication Date: 2021-07-27
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

[0003] In addition, the distinction and identification of species is the basis of various research work. However, the appearance of adults of giant clam oysters and other species of giant clam species is relatively similar, and it is difficult to directly identify them through morphological characteristics, especially in the early stages of development. difficulty

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  • A kind of identification primer and method suitable for giant clam oyster, common species of giant clam in the South China Sea and its hybrids

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Embodiment 1

[0017] see figure 1 , using phenol-chloroform extraction method to extract the genomic DNA of 40 individuals of giant clam oyster, scaleless giant clam, scaled giant clam, long giant clam, safflower giant clam and nova giant clam. The specific method is to cut the adductor muscle fully, absorb the water with clean filter paper, put it into a 1.5mL centrifuge tube, then add 400μl lysate and 10μl proteinase K (10mg / ml), mix well on the shaker, 55 Digest in a water bath at ℃ for 3-5 hours until the lysate is clear. Add an equal volume of saturated phenol (200 μL), chloroform / isoamyl alcohol (24:1) (200 μL) mixture and extract three times. Precipitate DNA with 1 mL of absolute ethanol, wash with 70% alcohol, dry at room temperature and dissolve in 100 μL of ultrapure water. The concentration and purity of DNA were detected by ultraviolet spectrophotometer, and the integrity of DNA was detected by 1% agarose gel electrophoresis. The genome DNAs of oyster, scaleless, scaled, long...

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Abstract

The invention discloses an identification primer and method suitable for giant clam oysters, common species of giant clam in the South China Sea and hybrids thereof. The identification primers are TC114 primer pair, and its forward and reverse primers are as follows: F: 5'-AACACTGATTGGGCTCTT-3'R: 5'-TTGACGACGATGACGATA-3'. The present invention screens out a pair of safflower clam microsatellite primers from 60 pairs of giant clam microsatellite primers, which can be used in giant clam and South China Sea common species (scaleless giant clam, scaled giant clam, long giant clam, safflower giant clam and nova giant clam) The microsatellite primers that are commonly used in giant clam) and can be distinguished from the common giant clam in the South China Sea according to the size of the amplified bands, and have clear band patterns, good repeatability, and high reliability: TC114 primer pair, and can be used in giant clam and South China Sea Individual identification of hybrid species among common giant clams also has potential application value.

Description

technical field [0001] The invention belongs to the shellfish molecular marker and species identification technology in the aquatic biology technology, and specifically relates to a method of using microsatellite primers to identify giant clam species and common species of giant clam in the South China Sea (scaleless giant clam, scaled giant clam, long clam, tomato The identification primers and methods of Tridacna red and Tridacna nova) and the potential application value of identifying hybrid individuals between them. Background technique [0002] Tridacna is a kind of tropical large marine coral reef benthic shellfish. Common giant clam species distributed in the South China Sea mainly include scaleless giant clam, scaly giant clam, long giant clam, saffron giant clam, and nova giant clam. and giant oysters of the genus Giant oyster. Tridacna is a species of high economic value. It is the material basis of tropical marine life and fishery economy, providing resources suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888
Inventor 马海涛喻子牛张跃环高红梅肖述
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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