title: GSM5776393: MM+DMSP-2; Sulfitobacter sp. D7; RNA-Seq
library_construction_protocol: RNA extraction was carried out using the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). For disruption of cell pellets we used the OmniLyse lysis kit (Claremontbio). The rest of the RNA extraction protocol was according to manufacturer's instruction. Library preparation was carried out according to the RNA-seq protocol developed by Avraham et al., 2016. DNA was removed using TURBO DNase (Ambion), RNA was fragmented and phosphorylated (at the 3'prime) using FastAP thermosensitive alkaline phosphatase (Thermo Scientific). RNA from each sample was ligated with unique RNA barcoded adaptors at the 3', ensuring the strandedness of each transcript, using T4 RNA ligase 1 (NEB). RNA samples were pooled and treated with RiboZero (Gram-Negative Bacteria) kit (Illumina, San Diego, CA, USA) following manufacturer's instructions in order to remove ribosomal RNA. Samples were reversed transcribed using AffinityScript RT Enzyme (Agilent) to form cDNA, and amplified by PCR.