Replicate A of asample taken on transect 6 at 102.9 m from the source, targeting 16S_Prokaryote
The amplicons were separately purified using solid phase reversible immobilization (SPRI) paramagnetic bead-based technology (AMPure XP beads, Beckman Coulter) with a Bead:DNA ratio of 0.7:1 (v/v). The purified indexed amplicons were normalized to equimolar concentration using SequalPrep (Thermo Fisher Scientific). The final library pool was subjected to a denaturation step using NaOH to ensure the presence of single stranded DNA (ssDNA) fragments for cluster generation. Thus, the library consisted of ssDNA fragments with sequencing adapters and indices. The library pool was diluted and used for loading on the MiSeq system for cluster generation and sequencing. Sequencing was performed at a final concentration of 8.0 pM and with a 20% PhiX control library spike-in (Illumina Inc.). For cluster generation and sequencing, a MiSeq Reagent Kit v3 (600-cycle) (IIllumina Inc.) was used. Cluster generation and sequencing were carried out under the control of the MiSeq-inherited MiSeq Control Software (MCS) v2.5.0.5.