Soil DNA indirect extraction method for evaluating diversity of plant root system microflora
A microbial community and plant root technology, applied in the field of indirect extraction of soil DNA, can solve the problems of difficult removal of humus and low DNA recovery rate in the purification step, and achieve the effects of reducing health damage, low cost and strong applicability
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Embodiment 1
[0036] (1) Add 0.5ml pre-cooled sterile water to 0.5g watermelon root soil sample and mix well, vortex for 10 minutes, then add 0.5ml homogenization buffer A, vortex for 10 minutes, low speed (250g) Centrifuge at room temperature for 10 min, collect the supernatant and transfer to another centrifuge bottle.
[0037] (2) Add 1ml of pre-cooled homogenization buffer B to wash and resuspend, vortex for 10 minutes, centrifuge at low speed (250g) for 10 minutes at room temperature, take the supernatant, combine the obtained supernatants, and centrifuge at 10000rmp for 30 minutes at room temperature to recover bacteria Somatic cells, discard the supernatant.
[0038](3) Add 1.5 ml of washing solution C, wash for 5 minutes, and centrifuge at 10,000 rpm for 30 minutes at room temperature to precipitate bacterial cells. Add 160ul lysozyme (50mg / ml) and 20ul proteinase K (20mg / ml) to the bacterial cells, bathe in water at 37°C for 30-40min, then add 122ul cell lysate D to the sample, an...
Embodiment 2
[0049] (1) Add 0.5ml pre-cooled sterile water to 0.5g strawberry root soil sample soil sample and mix well, vortex for 10 minutes, then add 0.5ml homogenization buffer A, vortex for 10 minutes, low speed (250g ) at room temperature for 10 min, and the supernatant was collected and transferred to another centrifuge bottle.
[0050] (2) Add 1ml of pre-cooled homogenization buffer B to wash and resuspend, vortex for 10 minutes, centrifuge at low speed (250g) for 10 minutes at room temperature, take the supernatant, combine the obtained supernatants, and centrifuge at 10000rmp for 30 minutes at room temperature to recover bacteria Somatic cells, discard the supernatant.
[0051] (3) Add 1.5 ml of washing solution C, wash for 5 minutes, and centrifuge at 10,000 rpm for 30 minutes at room temperature to precipitate bacterial cells. Add 160ul lysozyme (50mg / ml) and 20ul proteinase K (20mg / ml) to the bacterial cells, bathe in water at 37°C for 30-40min, then add 122ul cell lysate D t...
Embodiment 3
[0062] (1) Add 0.5ml of pre-cooled sterile water to 0.5g rice root soil sample and mix well, vortex for 10 minutes, then add 0.5ml homogenization buffer A, vortex for 10 minutes, low speed (250g) Centrifuge at room temperature for 10 min, collect the supernatant and transfer to another centrifuge bottle.
[0063] (2) Add 1ml of pre-cooled homogenization buffer B to wash and resuspend, vortex for 10 minutes, centrifuge at low speed (250g) for 10 minutes at room temperature, take the supernatant, combine the obtained supernatants, and centrifuge at 10000rmp for 30 minutes at room temperature to recover bacteria Somatic cells, discard the supernatant.
[0064] (3) Add 1.5 ml of washing solution C, wash for 5 minutes, and centrifuge at 10,000 rpm for 30 minutes at room temperature to precipitate bacterial cells. Add 160ul lysozyme (50mg / ml) and 20ul proteinase K (20mg / ml) to the bacterial cells, bathe in water at 37°C for 30-40min, then add 122ul cell lysate D to the sample, and ...
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