RNA-Seq of microbiome from Ircinia ramosa
21 samples from experimental setup were chosen to undergo RNA sequencing derived from three distinct genotype from seven different treatment types. Tissue samples homogenized with mortar and pestle under liquid nitrogen, lysed using FastPrep24 bead beater, lysing matrix E and TRIzol. RNA extraction was performed using PureLink RNA Minikit (Ambion), prokaryotic mRNA was retained using Poly(A)Purist MAG Kit, and ribosomal RNA was depleted using Ribo-Zero rRNA removal kit using 50% Bacteria and 50% Plant Leaf probes, before final mRNA purified using ethanol precipitation. Sequencing was eprformed at the Ramaciotti Centre for genomics, Sydney, Australia using a TruSeq Stranded mRNA kit and sequenced using Illumina NextSeq 500 platform, generating 2x150 bp reads.