16S rRNA amplicon sequencing of the soil samples: soil DI at week 0 replicate 2
Sequencing was performed on the Illumina MiSeq platform using the primers 27F and 357R. PCR conditions: 98 C for 5 min; 98 C for 10 s / 60 C for 30 s / 72 C for 30 s (30 cycles); 72 C for 5 min. PCR reaction (total 25 ?l); 12.5 ?l NEB Next High Fidelity Master Mix, 0.5 ?l of each primer, 1.5 ?l of 3% BSA, 1?g of template DNA. Sample purification: Agencourt AMPure XP extraction kit (Beckman Coulter Company), ratio of AMPure XP to PCR reaction is adapted to 0.8 to 1. Indexing PCR: 98 C for 5 min; 98 C for 10 s / 55 C for 30 s / 72 C for 30 s (8 cycles); 72 C for 5 min. Products are pooled equimolar to a final concentration of 4 nM and sequenced using the MiSeq Reagent kit v3 (600 cycles) for paired end sequencing.