Amplicon sequencing of metagenomic sample from terrestrial aquifer
All PCR reactions were conducted in triplicate with the appropriate annealing temperature as previously described (Vigneron et al., 2016). No amplification was observed with procedural DNA extraction and PCR negative controls. Replicate amplicons were pooled and purified from agarose gels using a Qiagen MinElute Gel purification kit (Qiagen, Hilden, Germany). PCR products were indexed using a Nextera XT kit (Illumina Inc., San Diego, CA, USA) and diluted to equimolar concentration according to the manufacturers recommendations. The DNA library was diluted to a concentration of 4 pM and sequenced using pair-end Illumina MiSeq sequencing. Sequencing was performed using an Illumina Miseq v3 kit (Illumina Inc.), as recommended by the manufacturer, to obtain 2x300bp pair-end sequences.