Illumina NovaSeq 6000 paired end sequencing; The SPOC domain is a phosphoserine binding module that bridges transcription machinery with co- and post-transcriptional regulators - RNAseq
The SPOC domain is a phosphoserine binding module that bridges transcription machinery with co- and post-transcriptional regulators - RNAseq
RNA-seq samples were prepared in three biological replicates. Cells were harvested and counted, 20% Drosophila S2 cells were added as a spike-in control.   Cell pellets were resuspended in 1 mL TRI reagent (Sigma). 200 µL Chloroform (Applichem) were added, samples were mixed and centrifuged at 4°C, max. speed for 15 min. The upper phase was transferred to a new tube and subjected to isopropanol precipitation. 20 µg of RNA were treated with 40 U DNase I (Roche) at 37°C for 30 min and purified by phenol-chloroform extraction and Ethanol precipitation. rRNA was depleted. DIDO and SHARP RNA-seq libraries were prepared with NEBNext rRNA depletion kit v2 (Human/Mouse/Rat) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs) according to the manufacturer's instructions. RBM15 RNA-seq libraries were prepared using Lexogen RiboCop rRNA Depletion kit HMR V2 and Lexogen CORALL Total RNA-seq Library Prep kit according to the manufacturer's instructions. 600 ng total RNA were used as an input for rRNA depletion.