Method for extracting genome DNA from phyllachora ascocarp

A genome and fungus technology, applied in the field of fungal DNA extraction, can solve the problem that the method of extracting DNA is rarely studied.

Inactive Publication Date: 2016-05-18
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the method of extracting DNA from the ascomycetes of M.

Method used

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Experimental program
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Embodiment Construction

[0018] The present invention will be further described below:

[0019] The present invention comprises the following steps:

[0020] (1) Use a sharp blade to remove 10-15 ascocarps from the host plant, without host tissue as much as possible, put them in a centrifuge tube, add a small amount of silicon dioxide and liquid nitrogen, and quickly grind them into powder;

[0021] (2) Add 800ul of CTAB-free buffer solution and mix quickly, place on ice for 10min; centrifuge at 9000r / min at 4°C for 5min, discard the supernatant, and repeat step (2) twice;

[0022] (3) Add 750ul CTAB extraction buffer preheated at 65°C, mix thoroughly and place in a water bath at 65°C for 1 hour;

[0023] (4) Take out the materials in the warm bath and place them at room temperature. After cooling to room temperature, add 750 μl of phenol-chloroform-isoamyl alcohol mixture in a ratio of 25:24:1, shake well for 3-5 minutes, and centrifuge at room temperature at 12000 rpm for 10 minutes;

[0024] (5) ...

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PUM

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Abstract

The invention discloses a method for extracting genome DNA from phyllachora ascocarp. Compared with the prior art, the method has the advantage that the phyllachora ascocarp is utilized as a material for directly extracting DNA. Before the extraction of DNA, a lysate buffer solution is utilized for processing twice, excessive colloids and polysaccharides are removed by virtue of a principle that DNA is low in solubility at a low temperature, finally an extracting solution is added to split cell membranes to extract DNA, and phenol-chloroform-isoamylol is added in the extraction process to remove redundant proteins through extraction. The method is applicable to fungi materials with high contents of colloids, polysaccharides and proteins, the yield and the quality of the extracted DNA are not obviously influenced by the existence and the content of impurities, and the guarantee is provided for the subsequent molecular research of phyllachora; and the method has popularization and use values.

Description

technical field [0001] The invention relates to a method for extracting fungal DNA, in particular to a method for extracting genomic DNA from the ascomycetes of the mole fungus. Background technique [0002] Obtaining high-quality total DNA is the premise and basis for molecular biology research. Many scholars have studied the DNA extraction methods of different groups of fungi. The current extraction methods for fungal DNA mainly include CTAB method (cetyltrimethylammonium bromide) and SDS method (sodium dodecylbenzenesulfonate). It is mainly through the artificial isolation and cultivation of fungi to obtain hyphae, and then extract DNA from the mycelia. For the extraction of DNA from hyphae, the above two methods are widely used. The DNA extraction methods of different organisms (plants, animals, microorganisms) are different; different types or different tissues of the same type have different separation methods because of their different cell structures and components...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 李晓玲李成云刘林杨静鄢进陆彭莞云杨亚琼刘立娜王慧王一王春梅杨雅婷
Owner YUNNAN AGRICULTURAL UNIVERSITY
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