characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2019-08-14T17:02:26Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-14T09:13:42Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104211098
characteristics.characteristic: External Id
characteristics.text: CHALMERS
characteristics.characteristic: INSDC center name
characteristics.text: 2019-08-14T17:02:26Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-14T09:13:42Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1870116
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5974:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: 24 to 26
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Each mouse received 250.000 Lewis lung carcinoma cells suspended in 100 ul PBS, injected subcutaneously at the left flank. RNA was extracted using the Trizol-Chloroform-Isopropanol method. Tissue, a metal bullet and 1 ml Trizol was added to cool 2 ml eppendorf tubes, and tissue was homogenized in a Qiagen Tissuelyser at 25 Hz for 2-3 x 1,5 min, and incubated 5 min at RT. 200 ul Chloroform was added to all samples and shaken. Samples were incubated for 3-5 min at RT to ensure dissociation of nucleic acids from protein complexes, and spun at 12,000 g for 15 min, 4 ºC. The upper clear aqueous phases (approx. 450 ul) were transferred to new PCR clean tubes. This is the RNA. RNA was precipitated with 0.45 ml isopropyl alcohol (1:1 reaction) at RT, and samples were vortexed for 5-10 sec and incubated 10 min at RT. Samples were spun at 12000 g, 15 min, 4 ºC, and placed in -20C cold blocks. The aqueous phases were poured out, and pellets were washed with 900uL (-20C chilled) 75% ethanol and spun at 8000 g, 5 min, 4ºC. This step was repeated once. Pellets were dried by placing tubes horizontally on a rack so that the tops “hung off” the edge of the rack and the air flow from the hood moved past the open tops of the tubes in a perpendicular manner. Tubes were left for 3-5 minutes until pellets had “just” dried and had NO remaining liquid. Pellets were dissolved in 15-30µl of Ultrapure RNase/DNase free water. Sequencing libraries were prepared from 1μg total RNA using the TruSeq stranded mRNA library preparation kit (Cat# RS- 122-2101/2102, Illumina Inc.) including polyA selection. The library preparation was performed according to the manufacturers’ protocol (#15031047). Library preparation using TruSeq stranded mRNA sample preparation kit is an accredited method.
characteristics.characteristic: description
characteristics.text: Lewis lung carcinoma
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_1001770
characteristics.characteristic: disease
characteristics.text: no access to running wheel
characteristics.characteristic: environmental history
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: muscle tissue
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0002385
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: C57BL/6
characteristics.characteristic: strain
characteristics.text: Sample 2
characteristics.characteristic: title