organism: Mus musculus
description: Protocols: Pancreatic islets (50-200) were extracted from homozygous Pax6 floxed mice which were also either hemizygous or null for Pdx1-CreERT one week after the completion of tamoxifen treatment by pancreatic distension and digestion with collagenase. Islets were incubated in RPMI media containing 11 mM glucose, 10% FCS, 100 IU/mL penicillin, 100 µg/mL streptomycin, at 5% CO2 and 37 C for 24 h. Transgenic mice were crossed to produce offspring which were all homozygous for floxed Pax6 alleles and either hemizygous or null for Pdx1-CreERT. All mice were injected with 5 daily intraperitoneal injections of tamoxifen (2 mg). RNA was extracted with TRIzol (Invitrogen) using the manufacturer's protocol, with the addition of glycogen as a co-precipitate. Resuspended RNA was treated with DNAse I (Sigma-Aldrich), which was then heat inactivated. Polyadenylated RNA was purified with oligo-dT beads and the library constructed using the TruSeq Stranded kit (Illumina). Illumina Universal Paired-end adapters were used, and custom barcoding sequences incorporated during the amplification rounds (15 cycles) for multiplexing.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2017-04-27T17:01:59Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2017-05-16T18:32:08Z
attributes.tag: External Id
attributes.value: SAMEA104031716
attributes.tag: INSDC center name
attributes.value: unspecified
attributes.tag: INSDC first public
attributes.value: 2017-04-27T17:01:59Z
attributes.tag: INSDC last update
attributes.value: 2017-05-16T18:32:08Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5708:9102
attributes.tag: age
attributes.value: 10
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: genotype
attributes.value: Pdx1-CreERT / Pax6 fl/fl
attributes.tag: organism part
attributes.value: islet of Langerhans
attributes.tag: phenotype
attributes.value: beta cell specific Pax6 knockout
attributes.tag: sample name
attributes.value: E-MTAB-5708:9102
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL/6