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
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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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