Mus musculus
RIP-Cre_LKB1_KO_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).
ENA-FIRST-PUBLIC
2014-10-25T17:01:04Z
ENA-LAST-UPDATE
2018-03-08T20:28:15Z
External Id
SAMEA2735913
INSDC center name
Imperial College London
INSDC first public
2014-10-25T17:01:04Z
INSDC last update
2018-03-08T20:28:15Z
INSDC status
public
Submitter Id
E-MTAB-2791:RIP_LKB1_KO_3
age
12 to 14 week
broker name
ArrayExpress
common name
house mouse
genetic modification
gene knock out
genotype
floxed LKB1 with RIP-Cre
organism part
islet of Langerhans
phenotype
LKB1 knockout
sample name
E-MTAB-2791:RIP_LKB1_KO_3
scientific_name
Mus musculus
strain
C57BL/6