title: Halobacterium salinarum NRC-1; growth curve point 37h; mid-log; treated with TEX; biological replicate B
design: Halobacterium salinarum NRC-1 were grown in complex media at 37 C, under light and constant agitation of 125 r.p.m.. Samples were taken at 3 different time points in a growth curve: middle of exponential phase (17h, OD600 @ 0.3, early-log phase), end of stationary phase (37h, OD600 @ 0.5, mid-log phase) and gas vesicles release phase (86h, OD600 @ 1.1, late-log phase). Reference samples were cultured under standard growth conditions (37 C, 225 rpm, constant light) at mid-log phase (OD600 @ 0.5). Total RNA extraction was performed using mirVana miRNA isolation kit (Ambion) from biological duplicates (A and B). DNA contamination was removed with treatment with Turbo DNAse (Ambion) and verified by PCR. Briefly, dRNA-seq was performed as following: TEX+ sample was treated with TEX (Epicentre TER51020) and TEX- was incubated only with buffer at 30 C for 60 min and purified with RNeasy MinElute Cleanup (Qiagen). Samples were treated with TAP (Epicentre), purified and quantified by Quant-iT RiboGreen RNA Assay (Invitrogen). For strand specific dRNA-seq library preparation, TruSeq Small RNA Sample Preparation (Illumina) was used. Sequencing was performed using Miseq Reagent Kit v2 300-cycles (Illumina). One may use the following sequences in order to trim the adapters from fastq files: Read 1: TGGAATTCTCGGGTGCCAAGGAACTCCAGTCAC; Read 2: GATCGTCGGACTGTAGAACTCTGAACGTGTAGA
identifiers.namespace: SUB3836185
identifiers.id: 37h_tex_plus_B