Extraction method of goose parvovirus DNA (deoxyribonucleic acid)

A technology of goose parvovirus and goose embryo, which is applied in the field of goose parvovirus DNA extraction, can solve the problems of expensive kits, inconvenient use, cumbersome process, etc., and achieve the effect of simple operation, easy operation and simple equipment

Inactive Publication Date: 2011-11-23
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The phenol and chloroform / isoamyl alcohol methods require many reagents, and the preparation is troublesome and the process is cumbersome
However, the kit is expensive, and there is no small amount of kit, the operation process is strict, and it is inconvenient to use

Method used

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  • Extraction method of goose parvovirus DNA (deoxyribonucleic acid)
  • Extraction method of goose parvovirus DNA (deoxyribonucleic acid)
  • Extraction method of goose parvovirus DNA (deoxyribonucleic acid)

Examples

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Effect test

example 1

[0020] Example 1, the extraction of goose parvovirus DNA in goose embryo allantoic fluid

[0021] Freeze goose embryo allantoic fluid containing goose parvovirus LH strain (isolation and identification of goose parvovirus LH strain Liu Yuzhuo; Li Yin; Wei Xuetao; Zhang Jingfeng Jiangsu Agricultural Science Journal 2009-10-31) at -20°C until completely frozen Take it out and let it melt at room temperature. After doing this three times, take chloroform (analytical pure AR, Nanjing Ningshi Chemical Reagent Co., Ltd., relative molecular mass 119.38, in line with: GB / 682-89, technical requirements: chloroform (CHCL3) content Not less than 99.0%, ethanol (C2H5OH) content (stabilizer) 0.3%-1.0%, density (g / ml) 1.471-1.484) added to the same amount of the above allantoic fluid, shake at room temperature for 20 minutes, 3500rpm Centrifuge for 10 minutes, absorb the supernatant, then add an equal volume of chloroform, oscillate, and centrifuge. After repeating this three times, absor...

example 2

[0023] Example 2, Electrophoresis results of PCR products in different layers of concentrated GPV-containing goose embryo allantoic fluid subjected to sucrose density gradient centrifugation

[0024] Goose embryo allantoic fluid containing goose parvovirus LH strain (isolation and identification of goose parvovirus LH strain Liu Yuzhuo; Li Yin; Wei Xuetao; Zhang Jingfeng Jiangsu Agricultural Science Journal 2009-10-31) was frozen and treated with chloroform according to the method of the present invention , absorb the supernatant and concentrate it with PEG, add it to the solution of different sucrose density gradients, centrifuge at 38000rpm for 4 hours, absorb the solution in layers and store it, absorb the solutions of each layer and put them into EP centrifuge tubes, boil for 5 minutes in boiling water, and centrifuge at 8000rpm for 5 minutes . Aspirate the supernatant for PCR determination, the results are shown in image 3 . Take the PCR electrophoresis strip laye...

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Abstract

The invention relates to an extraction method of goose parvovirus DNA (deoxyribonucleic acid), belonging to the field of biotechnology. The extraction method comprises the following steps: adding equal-volume of chloroform to a goose embryo allantoic fluid containing goose parvovirus, vibrating at room temperature for 15-20 minutes, centrifuging at 3500 rpm for 10 minutes, sucking supernatant, then adding equal-volume of chloroform, vibrating, centrifuging, repeating three times, sucking the supernatant, adding the supernatant into an EP (electropolished) tube, boiling in boiling water for 3-5 minutes, centrifuging at 8000-10000 rpm for 5 minutes, and sucking the supernatant to obtain the goose parvovirus DNA. In the extraction method provided by the invention, only chloroform is used as the reagent, and the virus is split to the allantoic fluid and fat and protein large particles are removed by repeated freezing and thawing, boiling and centrifuging; and the extraction method is convenient for operation, has low cost and high efficiency and is convenient for laboratory operation.

Description

technical field [0001] The invention relates to a method for extracting goose parvovirus DNA, belonging to the field of biotechnology. Background technique [0002] Trichloromethane (Trichloromethane) also known as chloroform, molecular formula is [CHCl 3 ]. A compound formed by replacing three hydrogen atoms in a methane molecule with chlorine. It is a colorless and transparent heavy liquid, very volatile, with a spicy and sweet taste and a special aroma. Miscible with ethanol, ether, benzene, petroleum ether, carbon tetrachloride, carbon disulfide and volatile oil, etc., slightly soluble in water (1ml dissolves in about 200ml water at 25°C). Its main functions are: 1. Chloroform is a raw material for organic synthesis, mainly used to produce Freon (F-21, F-22, F-23). In addition, it is also used in organic synthesis and anesthetics; solvents for fats, rubbers, resins, oils, waxes, phosphorus, iodine, and adhesive acrylics; extractants for penicillin, essential oils, al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12R1/93
Inventor 刘宇卓李银黄欣梅赵冬敏张敬峰
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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