Method for rapidly extracting plants sample DNA

An extraction method and plant technology, applied in the field of molecular biology, can solve the problems of many extraction steps, troublesome, complicated, etc., and achieve the effect of simple process, many operation steps and fast process

Inactive Publication Date: 2008-07-16
CENT LAB TIANJIN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to overcome the shortcomings of traditional sample DNA extraction steps,

Method used

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  • Method for rapidly extracting plants sample DNA
  • Method for rapidly extracting plants sample DNA
  • Method for rapidly extracting plants sample DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Using the rapeseed sold in the market as a sample, the method of the invention is used to rapidly extract DNA. The preparation method is as follows:

[0073] (1) Take about 100 mg of rapeseed, add a small amount of liquid nitrogen to a mortar, and grind it quickly. Add liquid nitrogen repeatedly 3 to 4 times to grind to powder, and put it into a sterilized centrifuge tube.

[0074] (2) Add 800 μL of CTAB DNA extraction buffer, vortex mix, place in a water bath at 65°C for 0.5 h, and centrifuge at 12,000 r / min for 5 min.

[0075] (3) Take the supernatant in a sterilized centrifuge tube, add 3mol / L sodium acetate solution 1 / 10 times the volume of the supernatant and 0.8 times the volume of isopropanol, centrifuge at 12000r / min for 5min, discard the supernatant .

[0076] (4) 100 μL of 10% Chelex-100 was added to the precipitate, and the precipitate was fully dissolved in the solution, centrifuged at 12000 r / min for 10 min, and the supernatant was taken and stored for P...

Embodiment 2

[0091] Taking corn grains from the farm of our hospital as samples, the method of the present invention was used to rapidly extract DNA.

[0092] (1) Take about 100 mg of corn in a mortar, add liquid nitrogen to grind to powder, and put it into a sterilized centrifuge tube;

[0093] (2) Add 800 μL cetyltrimethylammonium bromide extraction buffer, vortex mix, place in a water bath at 65°C for 40 minutes at a constant temperature, and centrifuge at 15,000 r / min for 8 minutes;

[0094] (3) Take the supernatant in a centrifuge tube, add 3mol / L sodium acetate solution 1 / 10 times the volume of the supernatant and 0.8 times the volume of isopropanol, mix well, centrifuge at 15000r / min for 8min, discard supernatant;

[0095] (4) Add 200 μL of 10% Chelex-100 solution to the precipitate, and fully dissolve the precipitate in the solution, centrifuge at 12,000 r / min for 10 min, take the supernatant, test the DNA, save it for PCR amplification.

[0096] PCR test:

[0097] 1. Primer seq...

Embodiment 3

[0110] Using the soybean callus collected in our laboratory as a sample, the method of the present invention was used to rapidly extract DNA.

[0111] (1) Take about 100 mg of the sample, add liquid nitrogen to the mortar, grind it to powder, and put it into a sterilized centrifuge tube;

[0112] (2) Add 600 μL cetyltrimethylammonium bromide extraction buffer, vortex mix, place in a water bath at 65°C for 40 minutes, centrifuge at 12,000 r / min for 10 minutes;

[0113] (3) Put the supernatant in a centrifuge tube, add 1 / 10 times the volume of 3mol / L sodium acetate solution and 0.8 times the volume of isopropanol, mix well, centrifuge at 12000r / min for 10min, discard the supernatant;

[0114] (4) Add 100 μL of 10% Chelex-100 solution to the precipitate, and fully dissolve the precipitate in the solution, centrifuge at 15000 r / min for 10 min, take the supernatant, save it, and use it for PCR amplification.

[0115] 1. Composition of PCR reaction system:

[0116] 10× PCR reactio...

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Abstract

The invention discloses an improved method for extracting a plant sample DNA rapidly, which grinds the plant sample to be tested to powder in liquid nitrogen, then a certain amount of CTAB extraction buffer solution is added; the supernatant liquid is taken to be arranged in a centrifuge tube after being processed, a 3mol/L sodium acetate solution with one tenth of the volume of the supernatant liquid and isopropanol with 0.8 time of the volume of the supernatant liquid are added and the mixture is mixed evenly, the centrifugation is carried out for 5 to 10min and the supernatant liquid is discarded; 10 percent of Chelex-100 solution is added in the sediment for centrifugation, the taken supernatant liquid is the sample DNA which can be directly used for PCR amplification. The DNA extraction method which is adopted by the invention can complete the whole process of sample DNA extraction within 60 minutes. The quality standard of the extracted DNA meets the requirements of follow-up PCR amplification and is more applicable to the purpose of the rapid extraction of the sample DNA in production and scientific research processes.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to a preparation method of template DNA. More specifically, an improved and rapid method for extracting DNA from plant samples. Background technique [0002] Molecular biology techniques have penetrated into various aspects of biology, medicine, botany, genetics and zoology. The quality and yield of DNA extraction are key steps that directly affect PCR amplification. Since the advent of PCR technology in 1985, this technology has been applied to various information fields of life sciences because of its high sensitivity, strong specificity, and easy operation. , short time required and other advantages have been widely used in a variety of detection fields. The first step of PCR detection technology is the preparation of template DNA, that is, the extraction of DNA in the sample, which directly affects the result of PCR reaction. Extraction and purification of DNA from sam...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C07H1/08C07H21/04
Inventor 王永兰青阔程奕张丽华朱珠赵新
Owner CENT LAB TIANJIN ACADEMY OF AGRI SCI
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