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Method for extraction plant DNA

A plant, plant extraction technology, applied in the field of DNA extraction

Inactive Publication Date: 2008-11-12
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the current plant DNA extraction methods are only effective for plant extraction materials containing a small amount of polysaccharides, and are basically ineffective for plants or parts with high polysaccharide content, and provide a method for extracting plant DNA

Method used

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  • Method for extraction plant DNA
  • Method for extraction plant DNA

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Experimental program
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specific Embodiment approach 1

[0009] Specific embodiment one: the present embodiment extracts plant DNA and carries out according to the following steps: one, the plant extract material is ground into powder with liquid nitrogen, then gets 0.15~0.25g ground powder and transfers in 1mL extraction buffer solution I at a temperature of 45 ± 1 Water bath at ℃ for 10 minutes, then centrifuge at 10,000g for 10 minutes; 2. Take the centrifuged sediment from step 1 and transfer it to 0.5-1mL extraction buffer II, bathe in water for 10 minutes at a temperature of 65±1℃, and then add 0.5- Centrifuge 1mL of chloroform at 12000g for 5min; 3. Take the centrifuged supernatant from step 2 and add an equal volume of isopropanol to the supernatant, mix well and centrifuge at 10000-12000g for 10min, then use , volume is 0.5mL ethanol washing centrifugation precipitation 2 times, then air room temperature drying, promptly obtains the DNA of plant extraction material; Wherein the extraction buffer I in step 1 is made of ethyle...

specific Embodiment approach 2

[0013] Embodiment 2: The difference between this embodiment and Embodiment 1 is that in Step 2, the centrifuge tube containing the extraction buffer II and the centrifuged sediment is inverted 3 to 10 times during the water bath process. Other steps and parameters are the same as those in Embodiment 1.

[0014] This embodiment can ensure uniform and sufficient reaction, and improve the quality of plant DNA extraction.

specific Embodiment approach 3

[0015] Specific embodiment three: the difference between this embodiment and specific embodiment one is: the temperature of the isopropyl alcohol added in step three is 0~4 ℃ (precooling). Other steps and parameters are the same as those in Embodiment 1.

[0016] This embodiment can effectively precipitate DNA and increase the yield of plant DNA.

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Abstract

The invention provides a method for extracting plant DNA, relating to a method for extracting DNA. The method solves the problems that the prior plant DNA extraction method is only effective to an extraction material of the plant containing a small amount of amylose, and is ineffective to the plant or parts containing high content of the amylose. The extraction method comprises the following steps of: firstly, grinding the plant extraction material into powder with liquid nitrogen; then, transferring the powder into an extraction buffer liquid I for water bath; and centrifuging the extraction buffer liquid I; secondly, taking and transferring centrifugal precipitate in the first step to an extraction buffer liquid II for water bath; and then adding chloroform into and centrifuging the extraction buffer liquid II; and thirdly, taking centrifugal supernatant fluid in the second step and adding isovolumetric isopropanol into the supernatant fluid; evenly mixing and centrifuging the mixture; washing with ethanol and drying the centrifugal precipitate to obtain the DNA of the plant extraction material. The method is a plant DNA extraction method with wide applications, and the amylose in the plant can be removed completely in the process of extracting DNA without any effect of content of the amylose of the plant extraction materials, thereby improving efficiency, purity and quality of DNA extraction.

Description

technical field [0001] The invention relates to a method for extracting DNA. Background technique [0002] DNA is one of the main objects of molecular biology research. In the process of molecular biology operations, the quality of genomic DNA is the key factor affecting its success or failure. For example, polysaccharides, polyphenols, and pigments can seriously affect the activity of DNA polymerase during PCR, and interfere with the combination of primers and templates, resulting in failure of amplification. [0003] Plants, especially some forest trees, contain a large amount of polysaccharides. Polysaccharides are the main interfering substances in the extraction of plant DNA. Because of their physical properties are very similar to nucleic acids, polysaccharides usually co-precipitate with DNA during the extraction process, and it is very difficult to separate them from DNA. , thus seriously affecting the quality of the extracted DNA. At present, in the process of ext...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C07H1/06
Inventor 王玉成刘桂丰李慧玉姜静杨传平
Owner NORTHEAST FORESTRY UNIVERSITY