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Identification method of fistulobalanus albicostatus and amphibalanus amphitrite as well as primer

A barnacle and white ridge technology, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. problems, to achieve the effect of improving accuracy and efficiency

Inactive Publication Date: 2010-02-24
XIAMEN UNIV
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Problems solved by technology

At present, the main method of barnacle classification is morphological classification (Liu Ruiyu; Ren Xianqiu. Chinese Zoology: Invertebrates Volume 42 [M]. Beijing: Science Press, 2007), but the same species of barnacles in different In the habitat of the barnacle, its shape changes with the environment, and the morphological difference between the larvae and adults of barnacles is relatively large, which has caused great difficulties in the classification and identification of barnacle samples and reduced the accuracy of identification.

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  • Identification method of fistulobalanus albicostatus and amphibalanus amphitrite as well as primer

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Embodiment Construction

[0046] 1) Extract the DNA of white-ridged barnacles and barnacles by conventional methods

[0047] Using the phenol-chloroform extraction method to extract the DNA of the white ridge tube barnacles and the grain barnacles, the specific steps are as follows:

[0048] (1) Take about 10 mg of adductor muscle from barnacles, put it into a 1.5 mL Eppendorf centrifuge tube with 200 μL of double distilled water, cut it into pieces with scissors, and let it stand for 0.5 h.

[0049] (2) Centrifuge at 10,000r / min for 1min, discard the supernatant.

[0050] (3) Add 564 μL of extraction buffer, 30 μL of 10% SDS (weight to volume ratio) solution to a final concentration of 5 mg / ml, add 6 μL of proteinase K (10 mg / ml) to a final concentration of 100 μg / ml, and mix well. Water bath at 55°C for 2 to 3 hours, turning over every 10 minutes during this period, until the tissue is completely digested.

[0051] (4) Cool the solution to room temperature, add 600 μL of phenol:chloroform:isoamyl a...

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Abstract

The invention relates to the identification of barnacles, in particular to an identification method of fistulobalanus albicostatus and amphibalanus amphitrite as well as a primer. The primer is a mixture primer of 18SCOM, 18SB1and 18SW1. The identification method comprises the following steps: extracting the DNA of the fistulobalanus albicostatus and the amphibalanus amphitrite; carrying out a PCRamplified reaction; making an electrophoretogram; and judging a result. The mixture primer of 18SCOM, 18SB1and 18SW1 is utilized for the PCR amplification of the DNA of the fistulobalanus albicostatus and the amphibalanus amphitrite which are identified by the electrophoretogram and can be effectively distinguished according to the lengths and the number of target fragments of an electrophoresisresult, and accordingly, the accuracy and the efficiency of the identification are improved.

Description

technical field [0001] The invention relates to the identification of barnacles, in particular to a method and primers for identifying white-ridged tube barnacles and barnacles. Background technique [0002] Barnacles belong to Arthropoda, Crustacea, Maxillopoda, Thecostraca, Cirripedia, Thoracoca, none Sessilia, Balanomorpha. A total of 541 species in 8 families have been recorded so far, including about 110 species in my country. [0003] Barnacles are common fouling organisms, mainly distributed in shallow water areas from the intertidal zone to the subtidal zone. If they attach to the bottom of a ship, it will greatly reduce the speed of the ship. It costs tens of billions of dollars to eliminate barnacles worldwide every year; Wakato If the pot is attached to the underwater pipeline system, it will easily cause blockage and accidents; in addition, barnacles can damage the paint protection layer of metal structures, and compete with mussels, oysters and other cultured ...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10
Inventor 王麟鹤梁翠王德祥
Owner XIAMEN UNIV