16s rRNA gene sequencing: roots of lettuce
Two replicated experimental aquaponics units  were used. Each individual aquaponics system was treated as an experimental unit and two units per treatment (control and Bacillus-Sanolife PRO-W). DNA was extracted from four samples of lettuce roots from four aquaponic systems (total of 16 roots).  After amplification and  purification, DNA was quantified using the PicoGreen assay (Invitrogen) and the quality was checked using a Bioanalyzer (Agilent). The Illumina MiSeq integrated next-generation sequencer (Illumina Inc., USA) and a MiSeq Reagent Kit v3 (600 cycles) (Illumina Inc. USA) were  used to sequence the prepared amplicon libraries. The Nextera XT adaptors were used to multiplex the amplicon libraries before loading onto the MiSeq and sequencing.