Kit and method for rapidly extracting plant genome DNA

A plant genome and kit technology, applied in the direction of recombinant DNA technology, DNA preparation, etc., can solve the problems of inability to guarantee the integrity of the genome, high price, low extraction volume, etc., achieve good amplification effect, fast operation, and avoid pollution Effect

Inactive Publication Date: 2015-03-11
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the DNA extraction kit contains the reagents and necessary consumables required for extracting plant DNA, the operation is simple, but it is expensive and the amount of extraction is small; the kits developed by some companies also use organic reagents such as chloroform, and organic solvents cannot be avoided pollution problem
Moreover, for example, the instruction manual of the whole-type gold rapid extraction plant gDNA reagent states that its amplification limit is 2000bp, indicating that it cannot guarantee the integrity of the genome, and the restriction effect on the PCR amplification of large fragment genes is more obvious

Method used

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  • Kit and method for rapidly extracting plant genome DNA
  • Kit and method for rapidly extracting plant genome DNA
  • Kit and method for rapidly extracting plant genome DNA

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Adopt the method of the present invention to extract the genomic DNA of different crops

[0048] At present, the approved commercial genetically modified crops include soybean, corn, rapeseed, sugar beet, potato, rice, tomato, cucumber, papaya and so on. As a rapid plant genome preparation method suitable for the detection of genetically modified products, we hope that this method can be universally applicable. Therefore, we have selected transgenic crops currently approved for commercialization as our detection objects.

[0049] Specific steps are as follows:

[0050] 1. Extraction of genomic DNA from different crops

[0051] (1) Leaves of transgenic soybeans, corn, rice, and potatoes were preserved in our laboratory; rapeseed, sugar beets, tomatoes, cucumbers, and papayas were purchased from Yonghui Supermarket. Take edible parts, put them into 2ml EP tubes, and add 2 capsules Put the sterilized small steel balls directly into liquid nitrogen. After the plant tiss...

Embodiment 2

[0074] Adopt the contrast of the genome DNA that method of the present invention extracts with laboratory common method

[0075] The gene amplified in Example 1 is a plant internal standard gene of about 360bp, so it has a better amplification effect. However, does the method of the present invention have its own advantages for larger exogenous genes? In order to compare the difference between this method and the CTAB extraction method, SDS extraction method, alkaline lysis extraction method and kit extraction method commonly used in the laboratory, the Os12g24050 gene transgenic rice was selected for detection in this example.

[0076] Specific steps are as follows:

[0077] 1. Using six different methods to extract genomic DNA from transgenic rice

[0078] Genome was extracted according to the method of the present invention in Example 1, CTAB extraction method, SDS extraction method, alkaline lysis extraction method, DNA extraction kit from Biomega Company and DNA extrac...

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Abstract

The invention belongs to the technical field of gene engineering, and develops a universal type kit used for rapidly extracting plant genome DNA for PCR amplification and a method, and the kit and the method are applicable to rapid extraction of DNA of multiple plant genomes, and satisfy research about plant molecular biology and genetics and rapid identification on transgenic crops. The kit comprises a balance solution, a lysate, a DNA combination solution, a DNA washing solution, a DNA eluent and a DNA adsorption column. The purpose of separating and purifying DNA is realized by utilizing the different combination degrees of a silicon matrix and DNA at different salt ion concentration and pH conditions. The method is rapid and simple in operation, is capable of finishing DNA extraction within 15 min, does not need phenol and chloroform extraction during extraction, and avoids pollution coming from organic solvents, and DNA extracted by employing the method has good amplification effect on 300-4500 bp plant endogenous gene and exogenous transgene.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and develops a general-purpose kit and method for rapidly extracting plant genome DNA suitable for PCR amplification by optimizing the traditional CTAB method for genome extraction, which can be used for various plant genomes The rapid extraction of DNA satisfies the research of plant molecular biology and genetics and the rapid identification of transgenic crops. Background technique [0002] Since 1994, when the world's first genetically modified plant product—storage-stable transgenic tomato was approved for commercial production in the United States and put on the market, genetically modified crops and genetically modified foods have traditionally improved crop stress resistance or food nutritional value and flavor. The unparalleled advantages of food quickly swept the world. According to the report of the International Service for the Application of Agricultural Biotechnology (I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 张煜隆杨靓吴祖建唐雅君刘小娟吴康承
Owner FUJIAN AGRI & FORESTRY UNIV
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