Analytical method for structures of microbial communities in composite filler in tower-type earthworm ecological filter
A microbial community and ecological filter technology, applied in the field of environmental microbial molecular ecology, can solve the problems of a wide variety of microorganisms, complex ecological filter fillings, and less research
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[0026] 1. Packing sample collection and pretreatment
[0027] Use a 10cm x 8cm sterile belt to collect about 50g of soil (10cm), sandstone (5cm) and crushed bluestone (3cm) fillers in the first, second and third stages of the tower earthworm ecological filter, and store them at -80°C after collection. Freeze-dried for 5 days. The freeze-dried filler samples were ground with a sterilized mortar, and then plugged with a 100-mesh stopper. After the plug was plugged, they were packed into new sterile belts and refrigerated at -20°C.
[0028] 2. DNA extraction and purification
[0029] Weigh 0.5g of the pretreated filler sample and add it to Bead Solution Tubes, then add 60ul of Solution S1 and 200ul of IRS solution and vortex for 15min on a vortex machine; transfer the supernatant from 10000g centrifugation for 30s to 2mlCollection Tubes, add 250ul of Solution Vortex S2 for 5s and then stand at 4°C for 5min; centrifuge at 10000g for 1min, transfer the supernatant to 2mlCollectio...
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