title: cell_culture_Hypoxia_HIF-2a_siRNA_rep1
organism: Homo sapiens
description: Protocols: MCF-7 cells were cultured in Dulbecco's modified Eagles medium DMEM (Invitrogen). Media were supplemented with and 10% Fetal Bovine Serum (FBS) (Sigma-Aldrich). Cells were incubated for 24 hours in an In vivo2 Hypoxia Work Station (Ruskin Technologies) in an atmosphere of either normoxia (21% oxygen) or hypoxia (1% oxygen). In addtion, cells were treated with scramble control, HIF-1a and/or HIF-2a siRNAs as describes in paper and cultured in hypoxia for 24 hours. Total RNA was prepared using the mirVana miRNA Isolation Kit (Ambion) and treated with DNaseI (TURBO DNA-free, Ambion). Ribosomal RNAs were depleted using RiboMinus Locked Nucleic Acid probes (Invitrogen). Poly A selected RNA-seq libraries were papered according to TruSeq RNA Sample Prep Kit (Illumina), directional PolyA selected RNA libraries were prepared using the ScriptSeq v2 RNA-Seq kit (Epicentre) and ribosomal depleted strand specific total RNA-seq libraries were prepared according to optimised protocol described in the paper.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-02-10T17:00:48Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:52:55Z
attributes.tag: External Id
attributes.value: SAMEA2242737
attributes.tag: INSDC center name
attributes.value: The Nuffield Department of Medicine, University of Oxford, Oxford, UK
attributes.tag: INSDC first public
attributes.value: 2014-02-10T17:00:48Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:52:55Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-1994:cell_culture_Hypoxia_HIF-2a_siRNA_rep1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: MCF-7
attributes.tag: common name
attributes.value: human
attributes.tag: sample name
attributes.value: E-MTAB-1994:cell_culture_Hypoxia_HIF-2a_siRNA_rep1
attributes.tag: scientific_name
attributes.value: Homo sapiens