Facsvantage se w diva cell sorter

A piece (~1cm³) of fresh Ren-PNG-Bali-16–03 was cut and homogenized gently in calcium-magnesium-free (CMF) artificial seawater (2 mL) using a sterile tissue homogenizer. The resulting suspension was passed through a 70 μm filter and the filtrate was used for cell sorting on a FACSVantage SE w/DiVa cell sorter (BD Biosciences, San Jose, CA USA). Particles were observed on the forward scatter (FSC) and side scatter (SSC) parameters, representing increasing 488 nm laser light scatter due to particle size and granularity, respectively. Eight gates were created along the FSC axis and particles from each were collected into separate tubes, resulting in partitions with increasing particle size composition (Supplementary Fig. 5). Metagenomic DNA was extracted from each using the Masterpure complete DNA extraction kit following manufacturer’s protocol (Epicenter, Madison, WI, USA). The DNA obtained was used for both 16S rRNA gene amplicon sequencing (1.5 K reads on average) and shotgun metagenomic sequencing (an average of 7 M single-end reads of 75 bps per sample) on an Illumina HiSeq 2500 platform as described above (Supplementary Table 1). 16S rRNA gene sequencing data were processed using Qiime as described above, and metagenomic data were assembled using SPAdes and processed as described below.