Rhizospheric community of a mine tailing established Acacia farnesiana on patch 1
Plant  roots were vortex-shaked and sonicated on sterile PBS solution, to get the rhizospheric pellet as described elsewhere (Lundberg et al. 2012). The metagenomic DNA was extracted from the pellets with the MoBio  PowerSoil extraction kit (MoBio Laboratories, Solana Beach, CA, USA).  PCR reactions were carried out to amplify the V3-V4 region (341F and 805R primers; 464 bp amplicon; Klindworth 2013) following the Illumina  MiSeq protocol with 5 overhangs. We performed triplicated PCR reactions for each sample, using the high fidelity Pfx platinum polymerase (Invitrogen, Thermo Fisher Scientific Corporation, Carlsbad, California, USA). Finally, amplicons from each replicate were pooled and purified with the Wizard SV Gel and PCR Cleanup System kit (Promega Corporation, Madison, Wisconsin, USA).