organism: Homo sapiens
description: 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.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-12-31
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-12-31
attributes.tag: External Id
attributes.value: SAMEA110174292
attributes.tag: INSDC center alias
attributes.value: The Francis Crick Institute
attributes.tag: INSDC center name
attributes.value: The Francis Crick Institute
attributes.tag: INSDC first public
attributes.value: 2022-12-31T00:58:23Z
attributes.tag: INSDC last update
attributes.value: 2022-12-31T00:58:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11854:ELAVL1_1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: HEK293
attributes.tag: cell_type
attributes.value: kidney cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: embryo stage
attributes.tag: disease
attributes.value: normal
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: kidney
attributes.tag: replicate
attributes.value: 1
attributes.tag: sample name
attributes.value: E-MTAB-11854:ELAVL1_1