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Process for extracting leukasmus rhabdovirus genome DNA of prawn

A white spot baculovirus and extraction method technology, applied to the field of shrimp white spot baculovirus genome DNA extraction, can solve the problems of troublesome operation and low yield of virus genome DNA extraction, and achieve convenient acquisition, easy grinding, and rapid homogenization while full effect

Inactive Publication Date: 2009-07-22
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 the supernatant, add proteinase K to a final concentration of 100ug / ml, add KCl to a final concentration of 50mmol / L, add pH 8.3-8.5 Tris·HCl to a final concentration of 10mmol / L, add gelatin to The final concentration is 0.1mg / ml, add NP-40 to a final concentration of 0.45%, add Toween-20 to a final concentration of 0.45%, add water to make up to 2ml. Mix the above mixture, boil in boiling water f...

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Abstract

The invention discloses a method for extracting genome DNA of prawn white spot baculovirus. It takes the surviving sick shrimp or fresh dead shrimp suffering from white spot baculovirus disease as samples, washes them with sterile water first, then puts the samples in a heating vessel, adds sterile water and immerses them to 2-3cm, and heats them. After boiling, keep for 8-10 minutes, add TN buffer and grind to make a homogenate, and process through medium-speed centrifugation and other steps to obtain WSSV genomic DNA. Because the present invention adopts the method of heating and boiling to pretreat the sample, the sample is easily ground, and the homogenate is rapid and sufficient; also because after the sample is heated, the animal protein, including the viral protein, coagulates and denatures, and the nucleocapsid of the virus particle is easy to separate from the tissue, The DNA yield is greatly improved; at the same time, for the early detection of diseased shrimp such as seedlings, whole shrimp can also be used, which facilitates the acquisition of samples. The invention provides a fast, convenient and effective way for extracting WSSV genome DNA.

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 the Taiwan area of ​​my country; in 1993, a large number of cultured shrimps in Japan and China died of white spot disease; at the end of 1994, Thailand, India, and Malaysia found more and more In 1995, white spot disease was reported in the United States; in 2001, white spot disease of cultured prawns occurred in Europe and Australia. 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 ...

Claims

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Application Information

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Patent Type & Authority Patents(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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