title: GSM2935038: TRMT2A_HEK293_miCLIP_rep_2; Homo sapiens; RIP-Seq
library_construction_protocol: For methylation-iCLIP experiments, TRMT2A expression constructs harbouring in-frame FLAG-epitope tags were used. The substrate-trapping point mutant form of TRMT2A had a glutamate residue in the catalytic domain (see Results) substituted with an alanine residue. Exponentially growing cells were transfected with cDNA constructs using Lipofectamine2000, and cells were harvested 24 hours later. Methylation-iCLIP was subsequently performed as described previously (Hussain et al. 2013a) using an anti-FLAG antibody to immunoprecipitate ectopically-expressed TRMT2A complexes. Prepared libraries containing in-line barcodes were sequenced on the Illumina HiSeq4000 platform. Cells were harvested by washing in Phosphate Buffered Saline (PBS) and then incubating with Trypsin-EDTA followed by further washing of detached cells in PBS.
attributes.tag: GEO Accession
attributes.value: GSM2935038