title: CT26 cell line rep2
organism: Mus musculus
description: 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.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-02-07T17:00:55Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T17:09:21Z
attributes.tag: External Id
attributes.value: SAMEA2343002
attributes.tag: INSDC center name
attributes.value: tron-mainz
attributes.tag: INSDC first public
attributes.value: 2014-02-07T17:00:55Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T17:09:21Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2292:CT26 cell line rep2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: CT26
attributes.tag: cell type
attributes.value: colorectal carcinoma cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: sample name
attributes.value: E-MTAB-2292:CT26 cell line rep2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: specimen with known storage state
attributes.value: fresh specimen