GSM7519494: m6AeCLIP-Seq-WT-eIF3d-Input-Tg-2; Homo sapiens; RIP-Seq
Cells were lysed in 50 mM HEPES-KOH pH 7.4, 150 mM KCl, 5 mM Mg(OAc)2, 1 % v/v NP-40, 0.25 mM DTT. For RNA-Seq, polyadenylated RNA was purified and libraries were made using NEBNext Ultra II RNA Library Prep Kit for Illumina. For m6A-eCLIP, libraries were constructed using the m6A-eCLIP kit from Eclipse Bio. For Subunit-Seq, eIF3d was immunoprecipitated using anti-HA beads, the cap-binding domain was isolated by TEV protease digestion, contaminating RNAs were removed by enzymatic treatment and libraries were made using the NEBNext Ultra II Directional RNA Library Prep Kit