SpyWTML_TAP_sRNA
Library preparation and sequencing performed at the Max Planck-Genome-Centre Cologne, Germany. rRNAs removal: Ribo-Zero rRNA Removal Kit (Illumina); TAP (tobacco acid phosphatase, Epicentre) treatment; T4 Polynucleotide kinase (Thermo Scientific); short (sample SpyWTML_TAP_sRNA) and long (sampleSpyWTES_TAP)RNAs fractions were separated with RNA Clean & Concentrator kit (Zymo Research); intermediate purification steps with RNA Clean & Concentrator kit (Zymo Research), Phenol:Chloroform:Isoamylalcohol (Roth) and Chloroform:Isoamylalcohol (Sigma). NEXTflex Small RNA Sequencing Kit v3 (Bioo Scientific) following manufacturers instructions until step G (22 cycles PCR). Libraries purification (substituting step H) with Agencourt AMPure XP beads (Beckman Coulter) following instructions from step I of the NEXTflex Rapid Directional qRNA-Seq Kit (Bioo Scientific). Quality control steps using Qubit (RNA BR Assay and dsDNA HS Assay) or bioanalyzer (RNA 6000 Nano Kit and High Sensivity DNA Kit).