title: GSM8170698: RIP_RF1_R2; Trypanosoma brucei; RIP-Seq
library_construction_protocol: UV crosslinked DRBD18-RNA, DRBD18(R→F)-RNA and DRBD18(R→K)-RNA complexes were immunopurified from the lysate using anti-MYC antibodies attached to MYC beads (GE Healthcare). The unbound fraction was also collected; 10% was used for RNA extraction and 2% for western blotting. Beads were washed three times with IP buffer (lysis buffer plus 150 mM NaCl) and treated with DNase 1 (Sigma) followed by proteinase K (Roche) after confirmation by western blot. RNA was extracted from beads with phenol/chloroform 16 and ethanol precipitated. RNA was isolated from the input fractions using Trizol and ethanol precipitation. Takara ThruPlex DNAseq kit (R400675) for Illumina. Samples were sequenced on a NovaSeq 6000, PE50, XP flow cell.