SAFB1 iCLIP in SHSY-5Y cells

We used iCLIP to investigate the RNA interactions of the RNA binding protein SAFB1 SH-SY5Y neuroblastoma cells were UV-crosslinked to mediate the formation of protein-RNA complexes. SAFB1 iCLIP was performed with partial RNase I digestion and following SDS-PAGE analysis of immunoprecipitated SAFB1 the cross-linked RNA was extracted, reverse-transcribed and PCR-amplified before being subjected to high-throughput sequencing using Illumina HiSeq 2500.