Primers and screening method of crassostrea hongkongensis triplet-base repetition microsatellite markers
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A technology of microsatellite markers and Hong Kong oysters, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve problems such as long breeding cycle, increase application prospects and scope, and death, and achieve PCR expansion Increase product stability, good polymorphism effect
Pending Publication Date: 2016-11-16
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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With the continuous expansion of the breeding scale, some serious problems have also appeared: since 1992, Hong Kong oysters along the coast of Guangdong have experienced periodic mass mortality; in addition, the breeding cycle of Hong Kong oysters is long, and the mortality rate in winter and spring is relatively high
In addition, in the application of microsatellite markers at this stage, it is mor...
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Embodiment 1
[0028] 1. Construction of Hong Kong oyster three-base repeat microsatellite DNA enrichment library
[0029] 1.1 Extraction and restriction digestion of genomic DNA
[0030] Take 30 mg of Hong Kong oyster adductor muscle tissue fixed with 70% alcohol, soak it in deionized water 3 times, and change the water after 15 minutes each time to completely remove the alcohol remaining in the muscle. Use clean filter paper to absorb water and put it into a 1.5mL centrifuge tube, then add 400μl lysate and 10μl proteinase K (10mg / ml), and digest in a water bath at 55°C for 3 to 5 hours. Add an equal volume of saturated phenol (200μL), chloroform / isoamyl alcohol (24:1) (200μL) mixture 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 to obtain the genomic DNA of Hong Kong oyster. UV spectrophotometer was used to detect DNA concentration and purity, and 1% agarose gel ele...
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Abstract
The invention discloses primers and a screening method of crassostrea hongkongensis triplet-base repetition microsatellite markers. By means of the screening method, a large quantity of crassostrea hongkongensis triplet-base repetition microsatellite markers can be obtained in a short time, and then an effective tool is provided for population genetic structure analyzing and molecular mark assisted breeding of the markers. According to the primers, amplification products of the primers have the good polymorphism, and the PCR amplification products are stable and can be applied to analysis on crassostrea hongkongensis diversity and a population genetic structure.
Description
Technical field [0001] The invention belongs to the field of microsatellite molecular markers of the Hong Kong oyster, and particularly relates to a primer and a screening method for the three-base repeat microsatellite marker of the Hong Kong oyster. Background technique [0002] The Hong Kong oyster Crassostrea hongkongensis, also known as the white oyster, belongs to the Mollusca, Bivalvia, Pterioidae, Ostridae, Crassostrea. Mainly distributed near the coastal estuaries of Guangxi, Guangdong and Hainan in China. Due to its large size, fat body, delicious taste, etc., its market value is much higher than other oysters. Therefore, it has been used in Guangdong Shantou, Shanwei, Huizhou, Zhuhai, Zhenhaiwan, Yangjiang, Zhanjiang, Guangxi Beihai, Qinzhou Bay and other areas. Widely farmed. As the scale of aquaculture continues to expand, some serious problems have also emerged: since 1992, Hong Kong oysters along the coast of Guangdong have been periodically experiencing large-sc...
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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 马海涛喻子牛肖述张跃环张扬向志明李军
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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