Aplysina cauliformis with host depletion
Sponge tissue (about 3-4 cm3) was collected from the sponge in situ (3 - 6 m depth, FL Keys, USA) and put in a sterile Whirl-Pak bag with seawater. Bagged samples were stored on ice in the boat until return to the laboratory where they were removed from the water and stored in a cryovial (no buffer) at -80 degrees C. Total DNA was extracted from the sponge tissue and either host depleted ('hd' in sample name) using a QIAmp DNA microbiome kit or not host depleted (DNA extracted with the Qiagen powersoil powerlyser kit) before use in metagenome sequencing. Metagenome library preparation was completed with TruSeq DNA PCR-Free LP Illumina prep kit and sent to Azenta Life Sciences for sequencing. DNA sequencing was performed on an Ilumina HiSeq 4000 using 150 nucleotide length reads with paired end sequencing.