Instrument: Illumina MiSeq
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Cells were then centrifuged, resuspended in 1 mL of buffer A (20 mMTris-HCl at pH 8.0, 150 mM KCl, 1 mM MgCl2, 1 mM DTT). Cells were then transferred to screw cap tubes containing 200 μL of glass beads (0.1 mm bead diameter; BioSpec). Cells were disrupted using Precellys®24 Homogenizer (three cycles: 6500 rpm for 20 s, followed by 1 min in ice; Bertin). The lysate was centrifuged at 17,000g for 30 min at 4 °C and the supernatant was applied to the column for affinity purification. MS2-affinity purification was performed as previously described (Lalaouna et al., 2017). Both lysates were applied on the same column (200 pmol of the MS2-MBP protein immobilized on an amylose resin). Afterward, RNA samples (output) were treated with TURBO™DNase (Ambion). cDNA libraries were prepared with ScriptSeq™ v3 RNA-Seq Library Preparation Kit (Illumina) ScriptSeq™