Soil DNA extracting method for evaluating diversity of microbial community of plant root system
A technology of microbial community and extraction method, applied in the field of extraction of microbial genomic DNA from root soil, can solve the problems of low DNA recovery rate, difficulty in removing humus in purification steps, etc., achieving low cost, reducing health damage, and applicability strong effect
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Embodiment 1
[0036] (1) Add 1.5 mL of sample soaking solution A to 0.5 g of watermelon root soil sample, vortex for 10 minutes, centrifuge at 10,000 rpm for 5 minutes, discard the supernatant, and repeat 3 times.
[0037] (2) Add 1.5 mL of sample immersion solution 2, place on ice for 5 minutes, vortex for 10 minutes, centrifuge at 10,000 rpm for 5 minutes, discard the supernatant, and repeat 3 times.
[0038] (3) Add 0.3 g of quartz sand and 1 glass bead with a diameter of 4 mm, and add 978 μl of phosphate buffer C (pH=8.0) to the sample. Vortex for 5 minutes.
[0039] (4) Add 122 μl of cell lysate D to the sample, and vortex for 5-20 minutes. Centrifuge at 13000rmp for 10min to precipitate debris.
[0040] (5) Transfer the supernatant to a clean centrifuge tube, add 250 μl of protein removal solution E to the tube, gently invert and mix 10 times by hand, leave at room temperature for 5 minutes, and centrifuge at 13,000 rpm for 5 minutes.
[0041] (6) Transfer the supernatant to a clea...
Embodiment 2
[0050] (1) Add 1.5mL sample soaking solution A to 0.5g strawberry root soil sample, vortex for 10 minutes, 10000 rpm, centrifuge for 5 minutes, discard the supernatant, repeat 3 times.
[0051] (2) Add 1.5 mL of sample immersion solution 2, place on ice for 5 minutes, vortex for 10 minutes, 10,000 rpm, centrifuge for 6 minutes, discard the supernatant, and repeat 3 times.
[0052] (3) Add 0.3 g of quartz sand and 1 glass bead with a diameter of 4 mm, and add 978 μl of phosphate buffer C (pH=8.0) to the sample. Vortex for 5 minutes.
[0053] (4) Add 122 μl of cell lysate D to the sample, and vortex for 5-20 minutes. Centrifuge at 13000rmp for 10min to precipitate debris.
[0054] (5) Transfer the supernatant to a clean centrifuge tube, add 250 μl of protein removal solution E to the tube, gently invert and mix 10 times by hand, leave at room temperature for 5 minutes, and centrifuge at 13,000 rpm for 5 minutes.
[0055] (6) Transfer the supernatant to a clean centrifuge tube...
Embodiment 3
[0064] (1) Add 1.5 mL of sample soaking solution A to 0.5 g of rice root soil sample, vortex for 10 minutes, 10,000 rpm, centrifuge for 5 minutes, discard the supernatant, and repeat 3 times.
[0065] (2) Add 1.5 mL of sample immersion solution 2, place on ice for 5 minutes, vortex for 10 minutes, 10,000 rpm, centrifuge for 7 minutes, discard the supernatant, and repeat 3 times.
[0066] (3) Add 0.3 g of quartz sand and 1 glass bead with a diameter of 4 mm, and add 978 μl of phosphate buffer C (pH=8.0) to the sample. Vortex for 5 minutes.
[0067] (4) Add 122 μl of cell lysate D to the sample, and vortex for 5-20 minutes. Centrifuge at 13000rmp for 10min to precipitate debris.
[0068] (5) Transfer the supernatant to a clean centrifuge tube, add 250 μl of protein removal solution E to the tube, gently invert and mix 10 times by hand, leave at room temperature for 5 minutes, and centrifuge at 13,000 rpm for 5 minutes.
[0069] (6) Transfer the supernatant to a clean centrifu...
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