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Genomic DNA extraction kit and method

An extraction method and genome technology, applied in the field of molecular biology experiments, can solve the problems of long experiment time, low efficiency, and easy pollution, and achieve the effects of simplified DNA extraction, low experiment cost, and reduced damage

Inactive Publication Date: 2019-04-02
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method needs to purchase liquid nitrogen in the process of genomic DNA extraction, and the laboratory without a grinder needs to use a mortar for grinding. The sample DNA is prone to cross-contamination, and phenol and chloroform organic solvents are harmful to human health.
The traditional extraction method takes a long time to experiment, high cost, low efficiency, easy to pollute, and has certain risks
At present, many biological companies have corresponding kits for different experimental samples, but most of them are expensive, and generally only suitable for a certain type of plants, fungi or bacteria, and the versatility is not strong

Method used

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  • Genomic DNA extraction kit and method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1 kit preparation

[0064] This embodiment provides a kit, including solution A, solution B, solution C, solution D, solution E and an adsorption column, wherein:

[0065] Solution A: 50mM Tris-HCl, 50mM EDTA, 10% SDS, 2M NaCl, sodium N-lauroyl sarcosinate 3g / 100mL;

[0066] Solution B: 3M NaAC, pH 5.2;

[0067] Solution C: isopropanol;

[0068] Solution D: 75% ethanol;

[0069] Solution E: ddH containing 50 μg / mL RNase 2 O.

[0070] It can be prepared according to the conventional solution preparation method.

Embodiment 2

[0071] The extraction of embodiment 2 plant genome DNA

[0072] (1) Take about 20-50mg leaves of rice, wheat, corn, cotton, pepper and tomato, cut them into pieces, and add them to a 1.5mL centrifuge tube;

[0073] (2) Add 400 μL of solution A;

[0074] (3) Heating in a water bath at 94°C for 10 minutes, and mixing by inverting every 3 minutes;

[0075] (4) After cooling to room temperature, centrifuge at 12,000 rpm for 1 min, and take 350 μL of the supernatant into a new centrifuge tube;

[0076] (5) Add 35 μL of solution B, mix by inverting, add 350 μL of solution C, mix by inverting, and place at room temperature for 2 minutes;

[0077] (6) Transfer 750 μL of the mixed solution to the adsorption column, centrifuge at 9000 rpm for 30 s, and discard the waste liquid in the collection tube;

[0078] (7) Add 650 μL solution D to the adsorption column, centrifuge at 9000rpm for 30s, discard the waste liquid in the collection tube, centrifuge again at 9000rpm for 30s, transfer...

Embodiment 3

[0080] The extraction of embodiment 3 fungi genome DNA

[0081] (1) Take 20-50mg mycelia of Fusarium graminearum, Penicillium oxalicum, Staphylococcus aureus and Penicillium chrysanthemi, add them to a 1.5mL centrifuge tube,

[0082] (2) Add 400 μL of solution A;

[0083] (3) Heating in a water bath at 94°C for 10 minutes, and mixing by inverting every 3 minutes;

[0084] (4) After cooling to room temperature, centrifuge at 12000rpm for 30s, and take 350μL of the supernatant into a new centrifuge tube;

[0085] (5) Add 35 μL of solution B, mix by inverting, add 350 μL of solution C, mix by inverting, and place at room temperature for 2 minutes;

[0086] (6) Transfer 750 μL of the mixed solution to the adsorption column, centrifuge at 9000 rpm for 30 s, and discard the waste liquid in the collection tube;

[0087] (7) Add 650 μL solution D to the adsorption column, centrifuge at 9000rpm for 30s, discard the waste liquid in the collection tube, centrifuge again at 9000rpm for...

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Abstract

The invention discloses a genomic DNA extraction kit comprising a solution A, NaAC, isopropanol, 75% ethanol and ddH2O containing 50 [mu]g / mL RNA enzyme. The solution A consists of Tris-HCl, EDTA, SDS, NaCl and N-sodium lauroyl sarcosine. The invention also discloses a method for extracting the genomic DNA by the kit. The kit and method provided by the invention are suitable for rapid extraction of general type genomic DNA from plants, fungi and bacterial samples, have no need of liquid nitrogen grinding, can effectively avoid sample contamination, have the advantages of low cost, short time,and no need of organic solvent extraction, and reduce the experimental dangerousness.

Description

technical field [0001] The invention belongs to the technical field of molecular biology experiments, and in particular relates to a kit and method for extracting genomic DNA, more specifically, a kit and method for extracting genomic DNA from plants, fungi and bacteria. Background technique [0002] With the rapid development of molecular biology, the extraction of genomic DNA and PCR technology are more and more widely used in biology-related research fields, such as molecular marker identification, mutant identification, genetic diversity analysis, gene cloning, etc., and genome The extraction efficiency of DNA directly affects the experimental process. The traditional genomic DNA extraction methods mainly include SDS method and CTAB method. First, the sample is broken by liquid nitrogen grinding, and the protein and DNA are separated by organic solvents such as phenol and chloroform, and finally precipitated with absolute ethanol or isopropanol. DNA. The traditional me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 王建华刘玉洋赵志勇杨俊花杨宪立
Owner SHANGHAI ACAD OF AGRI SCI
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