16s rRNA metagenome of Halichondria sitiens: replicate 1
DNA was extracted from bacterial fractions obtained from 1 cm3 fragment of sponge tissue using the Diatom DNA Prep kit (Galart Diagnosticum). DNA was extracted twice with phenol-chloroform and chloroform and precipitated in ethanol for the additional purification. V3-V4 region of 16S rRNA gene was amplified using standard degenerate primers fused with sequencing adapters (see Illumina guide for 16S Metagenomic Sequencing Library Preparation, Part # 15044223 Rev. B): Forward primer = CCTACGGGNGGCWGCAG, Reverse primer = GACTACHVGGGTATCTAATCC, adapter sequences are not shown.