organism: Homo sapiens
description: Protocols: COLO320DM cells were seeded and 24 hours later 1.8Gy irradiated. Subsequently, 24h later COLO320DM cells were treated with the secretome of 10x 1.8Gy irradiated CAFs or sham treated CAFs in 0.5% of serum. RNA was isolated 6 hours or 48 hours later. After trypsinization of COLO320DM, a solution of 5 x 106 cells was centrifuged (210 g, 5 min, 4°C) and cell pellets were washed twice with PBS. The supernatant was discarded and the cell pellet was stored at -80°C. RNA was isolated with the miRNeasy kit (Qiagen, Venlo, The Netherlands) Libraries for mRNA sequencing were prepared using the TruSeq stranded mRNA sample prep kit (Illumina). The starting material (100 ng) of total RNA was mRNA enriched using the oligodT bead system (Illumina). The isolated mRNA was subsequently fragmented using enzymatic fragmentation. Then first strand synthesis and second strand synthesis were performed and the double stranded cDNA was purified (Agencourt AMPure XP, Beckman Coulter). The cDNA was end repaired, 3' adenylated and Illumina sequencing adaptors ligated onto the fragments ends, and the library was purified (Agencourt AMPure XP). The mRNA stranded libraries were pre-amplified with PCR and purified (Agencourt AMPure XP). The libraries size distribution was validated and quality inspected on the 2100 Bioanalyzer
attributes.tag: ENA first public
attributes.value: 2017-11-01
attributes.tag: ENA last update
attributes.value: 2017-01-05
attributes.tag: External Id
attributes.value: SAMEA39868918
attributes.tag: INSDC center alias
attributes.value: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, Ghent, Belgium
attributes.tag: INSDC center name
attributes.value: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, Ghent, Belgium
attributes.tag: INSDC first public
attributes.value: 2017-11-01T17:02:37Z
attributes.tag: INSDC last update
attributes.value: 2017-01-05T15:41:30Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5364:Sample 5
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: COLO320DM
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: colorectal adenocarcinoma
attributes.tag: dose
attributes.value: 1.8
attributes.tag: irradiate
attributes.value: X-ray
attributes.tag: sample name
attributes.value: E-MTAB-5364:Sample 5