characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-02-01T17:03:23Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-06T12:01:57Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103891176
characteristics.characteristic: External Id
characteristics.text: UCL Institute of Child Health
characteristics.characteristic: INSDC center name
characteristics.text: 2018-02-01T17:03:23Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-06T12:01:57Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1580344
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5538:Sample 3 - Sox2CreERT2/+; ROSA26flox-stop-YFP/+
characteristics.characteristic: Submitter Id
characteristics.text: 8 weeks
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000034
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: Sox2CreERT2/+; Ctnnb1lox(ex3)/+; ROSA26flox-stop-YFP/+ and Sox2CreERT2/+; ROSA26flox-stop-YFP/+ mice were induced with a single intra-peritoneal injection of tamoxifen (Sigma) at a dose of 0.15 mg per gram of body weight at 4 weeks of age. 1 month post-induction mice were culled and Anterior pituitaries were dissociated mechanically into a single cell suspensions following incubation in an enzyme mix (0.5% w/v Collagenase type 2 (Lorne Laboratories Ltd), 0.1X Trypsin (Gibco) and 50 μg/ml DNaseI (Worthington) with 2.5 μg/ml Fungizone (Gibco) in Hank’s Balanced Salt Solution (HBSS (Gibco))) for 4 hours at 37°C. Cells were then washed in HBSS and suspended in PSB supplemented with 1% Fetal Calf Serum (PAA), 25mM HEPES and 1 ng/μl of propidium iodide (PI (BD Pharmigen)). Cells were flow sorted for YFP and PI fluorescence using a MoFlo XPD (Beckman Coulter, Fullerton, CA, USA) Flow Cytometer. Fluorescence was detected using a 530/540 filter, where PI positive cells were excluded and the YFP positive and negative fractions collected directly into RLT buffer (Qiagen) supplemented with 1% 2-mercaptoethanol (Sigma). The SMARTer v4 low input assay kit (Clontech) was used to create amplified cDNA using the strand-switching protocol. Flow sorted cells from Sox2CreERT2/+; Ctnnb1lox(ex3)/+; ROSA26flox-stop-YFP/+ and Sox2CreERT2/+; ROSA26flox-stop-YFP/+ mice were processed for total RNA extraction using the RNeasy Micro kit (Qiagen). cDNA library preparation was performed using 200pg of amplified cDNA with the Nextera XT preparation kit (12 cycles of PCR).
characteristics.characteristic: description
characteristics.text: Sox2CreERT2/+; ROSA26flox-stop-YFP/+
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: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: Sample 3 - Sox2CreERT2/+; ROSA26flox-stop-YFP/+
characteristics.characteristic: title