Method for extracting genome DNA of Jatropha curcas energy plant

An extraction method and energy plant technology, applied in the field of energy plant Jatropha jatropha genome DNA extraction, can solve problems such as inability to effectively remove

Inactive Publication Date: 2011-03-16
陈珂
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Problems solved by technology

[0002] In the traditional plant genome DNA extraction method, the CTAB (2%) method is mostly used, but Jatropha belongs to Euphorbiaceae and is a plant with a lot of milk. Even the leaves have a high milk content and contain a large amount of polysaccharides. and polyphenol components, the traditional cationic detergent hexadecane and trimethylammonium bromide (2% CTAB) method cannot effectively remove these impurities to obtain high-quality genomic DNA

Method used

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Embodiment

[0016] An energy plant Jatropha genomic DNA extraction method, comprising the following steps:

[0017] a) Extraction of DNA: first collect fresh young leaves of Jatropha jatropha, wash them with distilled water, and wipe off the water on the surface of the leaves; grind 0.1-0.2 grams of leaves into powder with liquid nitrogen in a mortar, and add about 10 mg when grinding Polyvinylpyrrolidone (PVP-40) to prevent oxidation; then transfer the ground powder to a 2mL centrifuge tube, add 0.5ml 65 ℃ preheated 4×CTAB extraction buffer (4%CTAB, 20mM EDTA, 3.5MNaCl, 100mM Tris-HCl (pH 8.0 and 2% β-mercaptoethanol), shake well, then place in a water bath at 65°C for 90 minutes, and shake slowly every 5 minutes to further mix the mixture thoroughly; after the mixture is cooled to room temperature, add an equal volume of Chloroform / isoamyl alcohol (24:1, V V -1 ) gently mixed and extracted, each extraction time was 5 minutes; after 8 minutes of high-speed centrifugation at 8000 rpm, th...

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Abstract

The invention relates to a method for extracting genome DNA, in particular to a method for extracting the genome DNA of a Jatropha curcas energy plant, which is characterized by comprising the following steps: a) extracting DNA; b) purifying DNA; c) storing the DNA at a temperature below -20 DEG C; and d) detecting the purity and concentration of the DNA under wavelengths of 260 nanometers, 280 nanometers and 230 nanometers by using ultraviolet spectrophotometer. The method for extracting the genome DNA of the Jatropha curcas energy plant can extract high-quality genome DNA of the Jatropha curcas energy plant and can be used in researches on population hereditary, phyletic evolution, molecular marker-assisted breeding and the like of Jatropha curcas.

Description

technical field [0001] The invention relates to a method for extracting genome DNA, in particular to a method for extracting genome DNA of energy plant Jatropha jatropha. Background technique [0002] In the traditional plant genome DNA extraction method, the CTAB (2%) method is mostly used, but Jatropha belongs to Euphorbiaceae and is a plant with a lot of milk. Even the leaves have a high milk content and contain a large amount of polysaccharides. and polyphenol components, the traditional cationic detergent hexadecane and trimethylammonium bromide (2% CTAB) method cannot effectively remove these impurities to obtain high-quality genomic DNA. Contents of the invention [0003] The present invention aims at the above technical problems and provides a genomic DNA extraction method of Jatropha Jatropha, an energy plant, which can effectively remove impurities and optimize the DNA extraction and purification process. [0004] Concrete technical scheme of the present inventi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C07H21/04C07H1/08
Inventor 陈珂尹春英胡尚连阮莉萍焦娟玉
Owner 陈珂
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