B1_S5_3.2
The total DNA was extracted from 1g of soil by Mannitol-PBS-CTAB method, adapted from Fatima et al. 2014. Humic acids were removed by adding 200 uL of HTR reagent from E.Z.N.A. Soil DNA Kit Protocol. The DNA was stored at -20 C. Two-step PCR of the v4 region was performed, First 30 cycles with broad range primers (Uni341F and Uni806R), then 15 cycles to add sequencing primers and adaptors. 250 bp Paired-end sequencing, of up to 192 samples, were performed on the Illumina MiSeq System using MiSeq Reagent Kits v2