title: MCF7 CTRL
organism: Homo sapiens
description: Protocols: MCF7 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with non-essential amino-acids (1%), penicillin and streptomycine (1%), gentamycin (0.1%), fungizone (0.1%) and 10% FCS. RNAs were purified from cultured cells using the High Pure RNA isolation kit (Roche). The integrity of RNA was checked with BioAnalyser 2100 (Agilent technologies) that indicated a RQI score >8 The libraries were prepared with Truseq mRNA Sample Prep kit (Illumina) from 1 microgram of total RNA following manufacturer's instructions. mRNAs were isolated by poly-A selection and fragmented (8 minutes at 94°). Fragmented mRNAs were used for reverse-transcription in the presence of Superscript II (Invitrogen) and random primers. After second strand synthesis, end-repair, A-tailing and purification, the double strand cDNA fragments were ligated to Truseq adapters containing the index sequences. Fifteen cycles of PCR in the presence of dedicated PCR primers and PCR master mix were applied to generate the final libraries.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2016-04-22T17:01:20Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T18:27:05Z
attributes.tag: External Id
attributes.value: SAMEA2731823
attributes.tag: INSDC center name
attributes.value: UNIVERSITY
attributes.tag: INSDC first public
attributes.value: 2016-04-22T17:01:20Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T18:27:05Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2663:MCF7 CTRL
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-2663:MCF7 CTRL
attributes.tag: scientific_name
attributes.value: Homo sapiens