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A kit and method for rapidly extracting plant genome dna

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

Inactive Publication Date: 2018-04-13
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A kit and method for rapidly extracting plant genome dna
  • A kit and method for rapidly extracting plant genome dna
  • A kit and method for rapidly extracting plant genome dna

Examples

Experimental program
Comparison scheme
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 tissue...

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 extracti...

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Abstract

The invention belongs to the technical field of genetic engineering. By optimizing the method for extracting genome by traditional CTAB method, a universal kit and method for rapidly extracting plant genome DNA suitable for PCR amplification are developed, 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. The composition of the kit includes: equilibration solution, lysis solution, DNA binding solution, DNA washing solution, DNA elution solution, and DNA adsorption column. The invention utilizes the different degrees of binding between the silicon substrate and the DNA under different salt ion concentrations and pH conditions to achieve the purpose of separating and purifying the DNA. The method is quick and simple to operate, and can complete the DNA extraction in 15 minutes. No phenol or chloroform extraction is required in the extraction process, and the pollution of organic solvents is avoided. The source-transformed gene has a good amplification effect.

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 Patents(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