Sponge Bacterial Community Profiling through 16S rRNA Sequencing

Genomic DNA from sponge tissue (≈0.3 g) for 16S rRNA gene amplicon sequencing was extracted using the Precellys© Evolution homogenizer with a Cryolys cooling unit (Bertin Technologies, Montigny-Le-Bretonneux, France) and a DNeasy PowerSoil Kit (Qiagen, Hilden, Germany), following the manufacturer’s instructions. Extractions were performed using both internal and external sponge tissue in order to obtain the whole bacterial community. DNA concentrations were measured using a Nanoquant Spectrophotometer (Tecan, Switzerland). Approximate 500 ng of DNA sample were sent to the Dalhousie University CGEB-IMR¹ for V4–V5 rRNA gene library preparation and sequencing. Primers used correspond to 517f GTGYCAGCMGCCGCGGTAA and 926r CCGYCAATTYMTTTRAGTTT (Walters et al., 2015 (link)). Samples were multiplexed using a dual indexing approach and sequenced using an Illumina Miseq with paired-end 300 + 300 bp reads. All PCR procedures, primers, and Illumina sequencing detail were as described in Comeau et al. (2017) . Sequences were deposited at NCBI as BioProject with accession ID PRJNA541486.