title: HeLa siControl 3
organism: Homo sapiens
description: Protocols: HeLa cells were cultured in Dulbecco's Modified Eagle Medium with GlutaMAX (Thermo Fisher) with 10% fetal bovine serum (Thermo Fisher). For siRNA-mediated depletion of eIF4A3, HeLa cells were transfected with Lipofectamine 2000 (Thermo Fisher) with 20 nM siRNA against human EIF4A3 or with a non-targetting control oligonucleotide. Cells were harvested 48 hrs after transfection, exposed to 140mJ/cm2 UV-C light and used for iCLIP. The iCLIP protocol was followed to isolate RNA bound to PRPF8 and amplify a cDNA library as described in the manuscript. An anti-PRPF8 antibody (A303-922A, Bethyl) was used for immunoprecipitation (reference doi:10.1038/nrg3141). Samples 2,3 and 4 for each condition were barcoded at the point of adapter ligation to the RNA, then multiplexed for the subsequent steps. Specific barcodes were used for each RT reaction, with random nucleotides also incorporated into cDNAs to distinguish between PCR duplication of cDNA products.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-10-14T17:02:27Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-10-04T14:40:02Z
attributes.tag: External Id
attributes.value: SAMEA4952633
attributes.tag: INSDC center name
attributes.value: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK
attributes.tag: INSDC first public
attributes.value: 2018-10-14T17:02:27Z
attributes.tag: INSDC last update
attributes.value: 2018-10-04T14:40:02Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7269:HeLa siControl 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: HeLa
attributes.tag: cell type
attributes.value: epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: cervical adenocarcinoma
attributes.tag: organism part
attributes.value: uterine cervix
attributes.tag: sample name
attributes.value: E-MTAB-7269:HeLa siControl 3
attributes.tag: scientific_name
attributes.value: Homo sapiens