title: Identification of transcriptome-wide mRNA targets of the ELAVL4 protein
design: PAR-CLIP from HEK293 cells over-expressing FLAG-HA-tagged ELAVL2 protein was performed as described previously (Hafner et al. 2010). Briefly, the growth medium of HEK293 cells was supplemented with 100 µM 4-thiouridine (4SU) for 16 hours prior to crosslinking. After decanting the growth medium, cells were irradiated uncovered with 0.15 J/cm2 of 365 nm UV light in a Stratalinker 2400. A 6 ml pellet was processed to whole lysate. FLAG-HA-tagged-ELAVL4 was immunoprecipitated with anti-FLAG antibody conjugated to protein G Dynabeads. The radiolabeled band corresponding to ~50 kDa ELAVL4-RNA complex was excised, the associated RNA isolated by phenol-chloroform extraction following proteinase K treatment, converted to a cDNA library and Illumina sequenced at the Rockefeller University Genomics Center. Note: (1) Submitted file is the raw sequencing file without any processing. (2) For information regarding how to process see PMID: 24860013, for a dataset collected and processed similarly.
identifiers.namespace: Rockefeller University
identifiers.id: ELAVL4_HEK293