characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-02-07T17:00:55Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T17:09:21Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2343004
characteristics.characteristic: External Id
characteristics.text: tron-mainz
characteristics.characteristic: INSDC center name
characteristics.text: 2014-02-07T17:00:55Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T17:09:21Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS400696
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2292:CT26 cell line rep3
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: CT26
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0006982
characteristics.characteristic: cell line
characteristics.text: colorectal carcinoma cell
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: CT26 colon carcinoma cell line was purchased in 2010 from the American Type Culture Collection (Product: ATCC CRL-2638, Lot Number: 58494154). Early (3rd, 4th) passages of cells were used for tumor experiments. Cells were routinely tested for mycoplasma. RNA from bulk CT26.WT cells was extracted in triplicate using a combination of Trizol (Invitrogen) and Qiagen RNeasy Micro kit. Barcoded mRNA-seq cDNA libraries were prepared in triplicate, from 5 ug of total RNA (modified Illumina mRNA-seq protocol) mRNA was isolated from total RNA using Seramag Oligo(dT) magnetic beads (Thermo Scientific) and fragmented using divalent cations and heat. Resulting fragments (160-220 bp) were converted to cDNA using random primers and SuperScriptII (Invitrogen) followed by second strand synthesis using DNA polymerase I and RNaseH. cDNA was end repaired, 5' phosphorylated and 3' adenylated according to the manufacturer's instructions. 3' single T-overhang Illumina multiplex specific adapters were ligated with T4 DNA ligase using a 10:1 molar ratio of adapter to cDNA insert. cDNA libraries were purified and size selected at 200-220 bp (E-Gel 2% SizeSelect gel, Invitrogen). Enrichment, adding of Illumina six base index and flow cell specific sequences was done by PCR using Phusion DNA polymerase (Finnzymes). All cleanups up to this step were done with 1,8x volume of Agencourt AMPure XP magnetic beads. All quality controls were done using Invitrogen's Qubit HS assay and fragment size was determined using Agilent's 2100 Bioanalyzer HS DNA assay.
characteristics.characteristic: description
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: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: CT26 cell line rep3
characteristics.characteristic: title