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Method for purifying genomic deoxyribonucleic acid (DNA) of plant seeds by using goldmag particles

A technology of gold magnetic particles and plant seeds, which is applied in the field of extracting genomic DNA from plant seeds, can solve the problems of inconvenient operation, low purification rate, and low purity in the background technology, and achieve shortened DNA extraction time, high DNA quality, and fast purification process Effect

Active Publication Date: 2012-02-08
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a simple and fast method for purifying genomic DNA of various plant seeds by using magnetic nanocomposite materials, so as to solve the problems of inconvenient operation, high cost, low purity and low purification rate in the background technology

Method used

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  • Method for purifying genomic deoxyribonucleic acid (DNA) of plant seeds by using goldmag particles
  • Method for purifying genomic deoxyribonucleic acid (DNA) of plant seeds by using goldmag particles

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1: the method for purifying genomic DNA from soybean with gold magnetic particles

[0039] 1) Lysis of plant samples

[0040] 1.1) Weigh soybeans greater than 100mg, grind them into fine powder with a mortar, then accurately weigh 100mg and place them in a 2ml centrifuge tube, add 300μl of lysate 1 [2%-5% polyvinylpyrrolidone (PVP), 2% ~5% cetyltrimethylammonium bromide (CTAB), 0.5%~2% β-mercaptoethanol, 0.005~0.01M ethylenediaminetetraacetic acid (EDTA), 0.5~2M NaCl, 0.05~0.2M Tris -HCl], vortex and mix well, put in a 65°C water bath for 25 minutes (invert the centrifuge tube once every 5 minutes during the period), take out the centrifuge tube, and let it stand at room temperature for 1 minute;

[0041] 1.2) Add 30 μl of 10 mg / ml RNase solution into the centrifuge tube, blow and aspirate to mix, put in a water bath at 37° C. for 5 min, and take out the centrifuge tube.

[0042] 2) Chloroform extraction

[0043] Add 0.7ml of chloroform to the centrifuge t...

Embodiment 2

[0053] Embodiment 2: the method for purifying genomic DNA from rapeseed with gold magnetic particle

[0054] 1) Lysis of plant samples

[0055] 1.1) Weigh more than 100mg of rapeseed, grind it into a fine powder with a mortar, then accurately weigh 100mg and place it in a 2ml centrifuge tube, add 200μl of lysate 1 [2%-5% polyvinylpyrrolidone (PVP), 2 %~5% cetyltrimethylammonium bromide (CTAB), 0.5%~2% β-mercaptoethanol, 0.005~0.01M ethylenediaminetetraacetic acid (EDTA), 0.5~2M NaCl, 0.05~0.2M Tris-HCl], vortex and mix well, put in a water bath at 65°C for 25 minutes (invert the centrifuge tube once every 5 minutes during the period), take out the centrifuge tube, and let it stand at room temperature for 1 minute;

[0056] 1.2) Add 40 μl of 10 mg / ml RNase solution into the centrifuge tube, blow and aspirate to mix, put in a water bath at 37° C. for 5 min, and take out the centrifuge tube.

[0057] 2) Chloroform extraction

[0058] Add 0.6ml of chloroform to the centrifuge t...

Embodiment 3

[0068] Embodiment 3: the method for purifying genomic DNA from corn with gold magnetic particle

[0069] 1) Lysis of plant samples

[0070] 1.1) Weigh more than 100mg of corn, grind it into a fine powder with a mortar, then accurately weigh 100mg and place it in a 2ml centrifuge tube, add 200μl of lysate 1 [2%-5% polyvinylpyrrolidone (PVP), 2% ~5% cetyltrimethylammonium bromide (CTAB), 0.5%~2% β-mercaptoethanol, 0.005~0.01M ethylenediaminetetraacetic acid (EDTA), 0.5~2M NaCl, 0.05~0.2M Tris -HCl], vortex and mix well, put in a water bath at 65°C for 30 minutes (invert the centrifuge tube once every 5 minutes during the period), take out the centrifuge tube, and let it stand at room temperature for 1 minute;

[0071] 1.2) Add 50 μl of 10 mg / ml RNase solution into the centrifuge tube, blow and aspirate to mix, put in a water bath at 37° C. for 5 min, and take out the centrifuge tube.

[0072] 2) Chloroform extraction

[0073] Add 0ml of chloroform to the centrifuge tube, shak...

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Abstract

The invention provides a method for simply and quickly purifying genomic deoxyribonucleic acid (DNA) of various plant seeds by using a magnetic nano composite material. The method comprises the following steps of: performing sample lysis, namely adding 200 to 800 microliters of PVP (poly vinyl pyrrolidone) with mass-to-volume ratio of 2 to 5 percent, 2 to 5 percent of CTAB (cetyl trimethyl ammonium bromide) and beta-mercaptoethanol with volume fraction of 0.5 to 2 percent into a plant seed sample; blending, and performing full lysis for 5 to 30 min in a water bath; and adding 10 to 50 microliters of 10 mg / ml RNase solution, and thus obtaining sample lysis solution with the genomic DNA. The lysis solution is suitable for all plant seeds. The obtained genomic DNA does not contain an inhibitor which inhibits a downstream polymerase chain reaction (PCR); and DNA has high quality, high yield and high integrity.

Description

technical field [0001] The invention relates to a method for extracting plant seed genome DNA. Background technique [0002] Since the advent of transgenic technology in the 1970s, it has been applied in the improvement of various crop varieties. Using this technology, traits can be transferred not only between varieties, but also between different families and genera; overcome the incompatibility of distant hybridization; improve crop quality; improve crop stress resistance; thus realize the transfer of multiple target traits . Practice has proved that this technology has broad application prospects in crop variety improvement and is an effective new breeding method. In the process of realizing this technology, the extraction and purification of plant seed genomic DNA are very critical. [0003] Nucleic acid purification methods can be roughly divided into two types, one is phenol chloroform extraction liquid phase purification method, and the other is solid phase purifi...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 崔亚丽魏旭旭晁旭张景阁赵稳操李谭杰
Owner XIAN GOLDMAG NANOBIOTECH
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