characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-10-25T17:01:04Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T20:29:50Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2735921
characteristics.characteristic: External Id
characteristics.text: Imperial College London
characteristics.characteristic: INSDC center name
characteristics.text: 2014-10-25T17:01:04Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T20:29:50Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS535451
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2791:RIP_AMPK_WT_3
characteristics.characteristic: Submitter Id
characteristics.text: 12 to 14 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: RIP-Cre_AMPK_WT_3 Protocols: Pancreatic islets (50-100) were extracted from Ins1LKB1KO mice or Ins1AMPKdKO and their wild type controls aging 12-14 weeks by pancreatic distension and digestion with collagenase. Islets were incubated in RPMI media containing 11 mM glucose, 10% FCS, 100 IU/mL penicillin, 100 ug/mL streptomycin, at 5% CO2 and 37 C for 24 h. RNA was extracted using RNAeasy kit according to the manufacturer's instructions (Qiagen) All libraries were prepared using the TruSeq RNA Sample Preparation Kits v2 (Illumina, San Diego, CA USA) according to the manufacturer's instructions. In brief, magnetic beads containing poly-dT molecules were first used to purify mRNA from 250 ng of total RNA. Second, samples were chemically fragmented and reverse transcribed into cDNA. Finally, end repair and A-base tailing was performed before Illumina adapters were ligated to the cDNA fragments. Purified samples were amplified by 15-cycle PCR. Amplified material was validated and quantified using an Agilent 2100 bioanalyzer and the DNA 1000 Nano Chip Kit (Agilent, Technologies, Santa Clara, CA, USA).
characteristics.characteristic: description
characteristics.text: gene knock out
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000506
characteristics.characteristic: genetic modification
characteristics.text: floxed AMPK without RIP-Cre
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: islet of Langerhans
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000006
characteristics.characteristic: organism part
characteristics.text: wild type
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: phenotype
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: C57BL/6
characteristics.characteristic: strain
characteristics.text: RIP_AMPK_WT_3
characteristics.characteristic: title