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Sample GSM1466809 Query DataSets for GSM1466809
Status Public on Aug 07, 2014
Title NC_rep2
Sample type SRA
 
Source name adipocytes
Organism Mus musculus
Characteristics transfection: control
Treatment protocol For transfection, primary preadiocytes were kept in culture medium until 80-90% confluence. Then Locked Nucleic Acid (LNAs) miRNA inhibitors (100nM) were transfected by Lipofectaimine RNAi Max (Invitrogen) according to the manufacturer’s instruction. 8 hours after transfection, cells were recovered in full culture media and induced to differentiation. Four days after differentiation RNAs were harvested for analysis.
Growth protocol Briefly, tissue depots were digested in 0.2% collagenase at 37ºC for 25-30min. Digested tissues were filtered through a 100um membrane and SVF cells were collected by centrifugation at 1500rpm for 5min. After centrifugation, the mature adipocytes floating on top were collected for genotying analysis. The freshly isolated SVF cells were seeded and cultured in DMEM containing 10% new born calf serum and 0.5% penicillin/streptomycin at 37ºC with 5% CO2. On confluence, the cells were induced to differentiate for 2 days with DMEM containing 10% FBS, 850nM of insulin, 0.5uM of dexamethasone (Sigma), 250uM of 3-isobutyl-methylxanthine (IBMX) (Sigma) and 1VM of rosiglitazone (Sigma) in DMEM. The induction medium was replaced with DMEM containing 10% FBS and 160nM insulin for 2 days. Then cell were incubated in DMEM with 10% FBS.
Extracted molecule total RNA
Extraction protocol Qiagen miRNeasy Mini Kit
NEBNext® Ultra™ Directional RNA Library Prep Kit
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina HiSeq 2000
 
Data processing Reads were aligned to mm10, using Tophat (version tophat-2.0.9) . Differential expression was performed using Cuffdiff (version 2.1.1).
Genome_build: mm10
Supplementary_files_format_and_content: RPKM and fold-change
 
Submission date Aug 06, 2014
Last update date May 15, 2019
Contact name Lei Sun
E-mail(s) [email protected]
Organization name Duke-NUS
Street address 8 College Road
City Singapore
ZIP/Postal code 169857
Country Singapore
 
Platform ID GPL13112
Series (1)
GSE60165 MicroRNAs are required for the feature maintenance and differentiation of brown adipocytes
Relations
BioSample SAMN02953563
SRA SRX670806

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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