Process for extracting leukasmus rhabdovirus genome DNA of prawn

A white spot baculovirus and extraction method technology, which is applied to the field of shrimp white spot baculovirus genomic DNA extraction, can solve the problems of troublesome operation, low yield of extracted viral genomic DNA, etc., and achieves convenient acquisition, easy grinding, and reduced requirements. Effect

Inactive Publication Date: 2007-07-18
SUZHOU UNIV
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Problems solved by technology

At present, the method of extracting the genomic DNA of the virus is to directly grind the fresh tissue at low temperature. Because the fresh animal tissue is difficult to homogenate, the operation is troublesome, and the yield of the viral genomic DNA is also low, which brings certain difficulties to the development of research work. influences

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  • Process for extracting leukasmus rhabdovirus genome DNA of prawn

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Embodiment

[0019] Take live shrimp or fresh dead shrimp suffering from white spot disease as samples for extracting WSSV-DNA, wash them with tap water, and then rinse them with sterile water; put the samples in a heating vessel, add sterile water to immerse 2.5cm, heat to boiling, and keep 10 minutes; take the whole shrimp from the juvenile shrimp, and take the abdominal muscle from the adult shrimp, weigh 5 grams of the sample, add weight Compare 10 times (W / V=mg / ml) TN buffer (50mmol / L Tris HCl, pH7.6, 0.4mol / L NaCl) was ground to make a homogenate; Centrifuge for 10 minutes, take 1ml of supernatant, add proteinase K to a final concentration of 100ug / ml, add KCl to a final concentration of 50mmol / L, add pH8.3-8.5 Tris·HCl to a final concentration of 10mmol / L, add gelatin To a final concentration of 0.1mg / ml, add NP-40 to a final concentration of 0.45%, add Toween-20 to a final concentration of 0.45%, and add water to make up to 2ml. Mix the above mixture, boil in boiling water for 1...

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Abstract

The present invention discloses process of extracting genome DNA of prawn white spot baculovirus. The process adopts disease prawn with white spot as sample, and includes the steps of washing disease prawn with sterilized water, boiling in sterilized water for 8-10 min, adding TN buffer solution and grinding into homogeneous slurry, centrifuging at medium speed, etc to obtain genome DNA of prawn white spot baculovirus. The process is fast, simple and effective.

Description

technical field [0001] The invention relates to a method for extracting genomic DNA of aquatic animal viruses, in particular to a method for extracting genomic DNA of prawn white spot baculovirus. Background technique [0002] At the end of the 1980s, large-scale deaths of cultured Penaeus monodon caused by the virus occurred in Taiwan in my country. In 1993, a large number of white spot disease occurred in cultured shrimps in Japan and China and died; Farmed prawns died of the disease; white spot disease was reported in the United States in 1995; white spot disease of cultured prawns occurred in Europe and Australia in 2001. So far, the disease has spread to the coastal countries of the world. The white spot disease of prawns has caused a devastating blow to the shrimp farming industry and caused immeasurable losses to the world shrimp farming industry. The prevention and treatment of the white spot disease of prawns has attracted people's attention. Research is one of the ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/33
Inventor 许雅香张茂友沈卫德朱玉芳李兵刘衬丽
Owner SUZHOU UNIV
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