characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-12-31T00:58:23Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-12-31T00:58:23Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110174292
characteristics.characteristic: External Id
characteristics.text: The Francis Crick Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2022-12-31T00:58:23Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-12-31T00:58:23Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12273106
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11854:ELAVL1_1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: HEK293
characteristics.characteristic: cell line
characteristics.text: kidney cell
characteristics.characteristic: cell type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: HEK 293 cells are grown to 90% confluency, then UV crosslinked 4x 150mJ/cm2 on ice. Afterwards, the crosslinked cells are scraped and pelleted, and kept at -80oC until the day of the experiment. The cell pellet is lysed in ice-cold iCLIP lysis buffer, and supplemented with 0.4U of RNase I per 1mg/ml lysate for RNA fragmentation. Protein G Dynabeads (Life Technologies) were coupled to rabbit anti-HuR antibody (from santa cruz), HuR protein-RNA complexes were isolated, and RNA was ligated to L3 adaptor on beads. The complexes were then visualised via infrared labelled adaptor, and purified with SDS-PAGE. cDNA was synthesized with Superscript IV Reverse Transcriptase (Life Technologies), and circularised by CircLigase II. Optionally, RecJ was used for adaptor removal, and Exonuclease I was used for RT primer removal.
characteristics.characteristic: description
characteristics.text: embryo stage
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: kidney
characteristics.characteristic: organism part
characteristics.text: 1
characteristics.characteristic: replicate
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: ELAVL1_1
characteristics.characteristic: title