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Method for extracting high-quality genome DNA from dry suriana maritime leaf and kit thereof

A technology of drying leaves, extraction methods, applied in the field of biology, which can solve the problems of poor DNA quality, failure of analysis results, and influence of experiments

Pending Publication Date: 2016-04-20
ANNOROAD GENE TECH BEIJING +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the molecular biology of Sea Rennenia, especially the extraction of genomic DNA from dried leaves of Sea Rennenia has not been reported, and the quality of the final DNA obtained when the conventional CTAB method is used to extract genomic DNA is generally poor. Affect the progress of subsequent tests, and may lead to the failure of the final analysis results in severe cases

Method used

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  • Method for extracting high-quality genome DNA from dry suriana maritime leaf and kit thereof
  • Method for extracting high-quality genome DNA from dry suriana maritime leaf and kit thereof
  • Method for extracting high-quality genome DNA from dry suriana maritime leaf and kit thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0062] Embodiment 1 The improved CTAB lysate of the present invention

[0063] PVP was added to the conventional CTAB lysate, and the ratio of each component was optimized to obtain an improved CTAB lysate.

[0064] The components of the improved lysate are shown in Table 1:

[0065] Table 1 CTAB lysate formula of the present invention

[0066]

Embodiment 2

[0067] Example 2 Extraction of Genomic DNA from Dry Leaf of Sea Mania

[0068] Using the CTAB lysate of Example 1 to lyse the dry leaf genomic DNA of the sea mantle

[0069] 1) Take about 100 mg of dried sea mantle leaf tissue, wash it with 80% ethanol prepared with sterile water, and cut the leaf into a size of about 1 mm;

[0070] 2) Put the cut leaves into the pre-cooled mortar, add liquid nitrogen to fully grind;

[0071] 3) Quickly transfer the ground tissue into an EP tube pre-filled with 800 μL of the CTAB lysate of Example 1 preheated at 70° C., mix well, and place the centrifuge tube in a 70° C. water bath for 30 minutes;

[0072] 4) After the water bath is completed and returned to room temperature, add 200 μL NaCl-TE, then add an equal volume of chloroform-isoamyl alcohol (24:1), mix thoroughly and centrifuge at 12,000 rpm for 10 minutes, and add the supernatant to a new EP tube middle.

[0073] 5) Add an equal volume of Tris saturated phenolbenzene-chloroform-is...

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Abstract

The invention provides a method for extracting genome DNA from a dry suriana maritime leaf and belongs to the biotechnology field. Specifically, the extraction method provided by the invention comprises the following steps: grinding and crushing dry leaf cells, extracting DNA by adopting a two-step process, removing impurities and purifying the DNA. By adopting the extraction method provided by the invention, secondary metabolism products, such as polyphenol and polysaccharide, rich in a dry tissue sample can be effectively removed, and finally the high-quality genome DNA is obtained. The DNA extraction method provided by the invention has the advantages that polyvinylpyrrolidone is added, polyphenol substances can be more effectively removed, and quality of the genome DNA is obviously improved. The DNA extraction method provided by the invention is applicable to extraction of genome DNA from dry leaves of trees such as suriana maritime.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a technology for separating and extracting plant genome DNA, in particular to a method for extracting genome DNA from dried leaves of sea renownia. Background technique [0002] The DNA in the cell mainly exists in the nucleus and is called genomic DNA. The extraction of plant gDNA is mainly used for gene library construction, PCR analysis and Southern hybridization, etc. gDNA extraction is the basic technology of plant molecular biology research. At present, a variety of methods have been developed to successfully extract gDNA from plant leaves, callus, tissue culture seedlings, fruits, phloem and other tissues and organs. [0003] However, in some cases, different plants or even the same plant tissue materials have different external properties such as sources, parts, and shapes, as well as internal characteristics such as chemical components and tissue structures. When extracting g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 魏少华韩改革玄兆伶李大为梁峻彬陈重建
Owner ANNOROAD GENE TECH BEIJING
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