title: metagenome of water sample: biofloc
design: Amplification of the full length 16S rRNA gene was performed using OneTaq 2X Master Mix. Universal 16S rRNA primers AGAGTTTGATYMTGGCTCAG (forward) and TACGGYTACCTTGTTACGACTT (reverse) containing Nanopore adapter at their 5 end was used (Frank et al., 2008). The PCR amplification profile was as follows; Initial denaturation: 94 C for 30 s, followed by 35 cycles of denaturation (94 C for 15 s), annealing (50 C for 15 s) and extension (68 C for 90 s). The PCR products were diluted (1:10) and used as template for index PCR reaction to incorporate Nanopore barcode. The indexed PCR products were then pooled, gel-extracted, and measured using Denovix high-sensitivity fluorescence quantification kit. Sequencing was performed using a Minion nanopore sequencer (Nanopore Technologies, Oxford, UK).