title: GSM7147805: K562, siMETTL3 KD, Input, r1, CLIP-seq; Homo sapiens; OTHER
library_construction_protocol: Cells were then harvested by centrifugation, lysed, and treated wiht 4 U/mL RNase I and 4 U/mL Turbo DNase at 37 °C for 5 min. Next, the cell lysate was cleared by centrifugation at 15000 g, 4 °C for 15 min, and kept on ice for the subsequent immunoprecipitation (IP). RBFOX2 antibodies were pre-conjugated to Protein A/G Magnetic Beads by incubation at room temperature for 45 min. The cell lysate was then added to the pre-conjugated beads and incubated at 4 °C overnight. RBFOX2-bound RNAs were end-repaired with T4 PNK, released by proteinase K treatment, and recovered. Libraries were prepared using NEBNext Small RNA Library Prep Set.