Identification primer suitable for hippopus and common tridacna varieties in South China Sea and hybrids thereof and method thereof

A technology for hybrids and Tridacna, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as difficulty in identification of morphological characteristics and identification in the early stage of development, and achieve intuitive and accurate results. Simple method and low cost effect

Active Publication Date: 2020-02-07
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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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 g

Method used

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  • Identification primer suitable for hippopus and common tridacna varieties in South China Sea and hybrids thereof and method thereof

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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 suitable for hippopus and common tridacna varieties in South China Sea and hybrids thereof and a method thereof. The identification primer is a TC114 primer pair, and a forward primer and a reverse primer of the identification primer are as follows: F: 5'-AACACTGATTGGGCTCTT-3'R: 5'-TTGACGACGATGACGATA-3'. A pair of microsatellite primer TC114 primerpair is screened from 60 pairs of tridacna crocea microsatellite primers, can be universally used in hippopus and common tridacna varieties (tridacna derasa, tridacna squamosa, tridacna maxima, tridacna crocea and tridacna ovalvata) in South China, and can be distinguished according to the size of an amplified band, meanwhile, the microsatellite primer TC114 primer pair is clear in banding pattern, good in repeatability and high in reliability, and has potential application value in individual identification of hybrids between hippopus and common tridacna in South China Sea.

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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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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