Metagenome of Scophthalmus maximus: intestine
DNA was isolated from intestines of fish sampled at 24 h or 30 days post-treatment with OTC dissolved in DMSO or with DMSO alone, using the DNeasy Power Soil Pro Kit (Qiagen) following the manufacturers instructions. Total DNA was assessed for its quality and quantity by 1.0% agarose gel electrophoresis, quantified with the dsDNA Quant-iT PicoGreen kit (Invitrogen) using a Victor3 fluorometer (PerkinElmer) and purity was measured with a NanoDrop instrument (Thermo Fisher Scientific). All samples passed the quality control and were subjected to 16S rRNA V3-V4 amplicon library construction. Briefly, the V3-V4 region of the 16S rRNA gene (~460 bp) was amplified using universal prokaryotic-specific primers as described by Klindworth et al. (2013). After purification of the amplicons with AMPure XP beads, a 16S rRNA gene metabarcoding library was prepared using the Nextera XT Index Kit (Illumina). Paired-end sequencing (2  300 bp) was subsequently performed on an Illumina MiSeq platform by Macrogen Inc. (Seoul, Republic of Korea).