characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-10-01T00:27:18Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-10-01T00:27:18Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA111476216
characteristics.characteristic: External Id
characteristics.text: Co-author
characteristics.characteristic: INSDC center name
characteristics.text: 2023-10-01T00:27:18Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-10-01T00:27:18Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS13570252
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12299:Sample 5
characteristics.characteristic: Submitter Id
characteristics.text: 80
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: The scWAT was removed from the inguinal region, snap frozen and stored at -80°C until further analysis RNA extraction from mice scWAT was performed by homogenisation in RNA STAT-60. Samples were broken up using a pestle and mortar under liquid nitrogen. The broken pieces were then put in 2 mL screw-top tubes with Lysing Matrix D (MP Biomedicals). 1 mL STAT-60 was added to the tissue samples, followed by homogenisation using Precellys 24 Touch. The homogenate was then transferred to 1.5 mL eppendorfs. Tubes were spun at 13,000 rpm for 5 min at 4°C to remove debris. The supernatant was removed and transferred to fresh tubes containing 200 µL of Chloroform. Samples were vortexed at maximum speed for 15 s, and spun at 12,000 g for 15 min at 4°C. A clear aqueous supernatant of around 600 µL for every 1 mL of STAT-60 added will have formed. The aqueous phase was transferred to fresh tubes containing 500 µL of isopropanol. Tubes were mixed by inversion several times and left at -20°C overnight. After overnight incubation, the precipitates were spun at 12,000 g for 10 min at 4°C to pellet RNA. After removing the supernatant, 1 mL of cold 70% ethanol (stored at -20°C) was added to the pellet and washed by vortexing briefly. Samples were then spun at 8000 g for 5 min at 4°C before discarding the supernatant. Pellets were air-dried for 10-15min on ice until transparent. Nuclease-free water was added to the pellet and incubated at 60°C for 10 min. Samples were placed on ice and re-suspended by pipetting up and down. 1 μg of total RNA was quality checked (RIN >7) using an Agilent Bioanalyser 2100 system and then used to construct barcoded sequencing libraries with Illumina’s TruSeq Stranded mRNA Library Prep Kit following manufacturer’s instructions
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: normoxia
characteristics.characteristic: environmental stress
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: inguinal fat pad
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: 129Ev/Sv
characteristics.characteristic: strain
characteristics.text: Sample 5
characteristics.characteristic: title