Plant genomes deoxyribonucleic acid (DNA) quick extraction device and testing method thereof

A plant genome and extraction device technology, applied in the field of plant genome DNA rapid extraction devices, can solve problems such as inconvenience and clumsy instruments, and achieve the effects of simple operation, simple judgment method, and simple and convenient extraction process

Inactive Publication Date: 2013-06-05
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No matter which method is used, a lot of expensive and clumsy instruments are used, such as centrifuges, water baths, etc., which brings a lot of inconvenience to the field testing

Method used

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  • Plant genomes deoxyribonucleic acid (DNA) quick extraction device and testing method thereof
  • Plant genomes deoxyribonucleic acid (DNA) quick extraction device and testing method thereof
  • Plant genomes deoxyribonucleic acid (DNA) quick extraction device and testing method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1.1 Plant Genomic DNA Rapid Extraction Device

[0032] According to the requirements of field testing, the air pressure method is used instead of the centrifuge; the method of matching the appropriate lysate is used instead of the water bath; the silica membrane is combined with the nucleic acid to obtain pure DNA.

[0033] The structural schematic diagram of the plant genomic DNA rapid extraction device of the present embodiment is as follows figure 1As shown, the plant genomic DNA rapid extraction device is composed of several syringes with sponges at the bottom, extraction columns 1 and several centrifuge tubes 2; the upper end 3 of the extraction column can be sealed and connected with the injection port of the syringe, and the extraction The lower end 10 of the column can be connected to the centrifuge tube 2, and the lower end 10 of the extraction column is connected with a loading cover 7, and the loading cover 7 is loaded with a silicon dioxide film 8 and then s...

Embodiment 2

[0056] Embodiment 2, the broad-spectrum applicability research of extraction method

[0057] Step 1: Plant material and plant genomic DNA extraction

[0058] Conventional crop seed and leaf material: corn, soybean, cotton, canola, rice, wheat seed and leaf.

[0059] Extraction and purification of plant genomic DNA: use the invented device to extract and purify plant genomic DNA according to the aforementioned steps. In addition, 5 ul of the extracted DNA solution was electrophoresed on a 1% agarose gel, and the quality of the DNA was judged according to its brightness and degree of diffusion; the concentration and purity of the extracted DNA were determined by an ultraviolet spectrophotometer NANO drop1000.

[0060] Step Two: Result Analysis

[0061] It can be seen from Table 1 that the device of the present invention is suitable for six common crops, can be used for seed and leaf materials, and has a very good broad spectrum. At the same time, the quality of the extracted...

Embodiment 3

[0067] Embodiment 3, the sensitivity verification of detection method

[0068] Step 1: Plant material and plant genomic DNA extraction

[0069] Genetically modified plant leaf material: genetically modified soybean GTS40-3-2 and MON89788.

[0070] Step 1: Plant Genomic DNA Extraction and Purification: Use the plant genomic DNA rapid extraction device of the present invention to extract and purify plant genomic DNA according to the aforementioned steps.

[0071] The second step: LAMP primer design, using the online primer design software Primer Explorer V4 ( http: / / primerexplorer.jp / elamp4.0.0 / index.html )

[0072] The third step: establishment of LAMP detection system, optimization of amplification system and amplification program.

[0073] The genome concentrations of genetically modified soybean GTS40-3-2 and MON89788 were diluted to 10 ng / μL and 1 ng / μL as templates, the concentration of primers was adjusted from 0.1 μM to 2 μM, and the constant temperature of LAMP wa...

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Abstract

The invention relates to a plant genomes deoxyribonucleic acid (DNA) quick extraction device and a testing method thereof. The plant genomes DNA quick extraction device is composed of a plurality of injectors with sponges on the bottoms, extraction pillars, and a plurality of centrifugal tubes, wherein the upper end portion of each of the extraction pillars can be connected with an injection port of each of the injectors in a sealing mode, the lower end portion of each of the extraction pillars can be connected with each of the centrifugal tubes, a loading seal cover is connected on the end portion of each of the extraction pillars, the loading seal cover is loaded with a silica film and is embedded in the inner side of the lower end portion of each of the extraction pillars in a clamping mode, and a through hole is formed in the center of the loading seal cover. According to the plant genomes DNA quick extraction device and the testing method thereof, compared with the prior art, large instruments of a centrifugal machine and the like are replaced by an air pressure method, the silica film is used for combining nucleic acid molecules, the plant genomes DNA quick extraction device is simple and effective, an extraction process is simple and convenient without any dependency of valuable instruments, testing time is short, the testing method is high in sensitivity, and the problem of quick testing of transgenetic products on site is resolved.

Description

technical field [0001] The invention relates to the extraction and detection technology of transgenic product nucleic acid in the technical field of bioengineering, in particular to a plant genome DNA rapid extraction device and a detection method thereof. Background technique [0002] With the rapid development of modern biotechnology, the application of genetic engineering technology in the field of agricultural production has become more extensive. Since 1996, when transgenic crops were first approved for commercial planting, their planting area has continued to grow at a rate of 8%. At present, it has reached 160 million hectares, an increase of 94 times compared to 1996. The top ten countries with more than one million hectares each. According to statistics, a total of 29 countries have planted genetically modified crops, of which 19 developing countries have planted almost half of the total area. The percentage increase in cultivated area in developing countries (11%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12Q1/68
Inventor 张大兵杨立桃张苗
Owner SHANGHAI JIAO TONG UNIV
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