characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-11-01T00:24:12Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-11-01T00:24:12Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112483898
characteristics.characteristic: External Id
characteristics.text: Max Planck Institute for Biology of Ageing
characteristics.characteristic: INSDC center name
characteristics.text: 2023-11-01T00:24:12Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-11-01T00:24:12Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14601248
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12579:YoungOld_Liver_tissue
characteristics.characteristic: Submitter Id
characteristics.text: mixed 3 and 18
characteristics.unit: month
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: hepatocyte
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Livers were dissociated using the Miltenyi liver perfusion kit (130-128-030) following the manufacturer’s instructions. For the sc-RNA sequencing experiment the isolated hepatocytes were washed two times with staining buffer (1x PBS, 2mM EDTA, 0.5%BSA) and 1 million hepatocytes were stained with Hoechst (15μg/ml) and Reserpine (5uM) for 30min at 37°C. Dead cells were excluded with PI staining (1μg/ml). Cells were sorted using a BD FACSARIA IIIU or Fusion Cytometer and 130um nozzle. The data were analysed using the BD FACSDiva and FlowJo softwares. part of nucleic acid library construction protocol scRNA-seq libraries were generated using an early version of the Smart-seq3xpress protocol. Reaction conditions were identical to the published final protocol deposited on protocols.io (https://www.protocols.io/view/smart-seq3xpress-yxmvmk1yng3p/v2). Briefly, cells were sorted into 300 nl of lysis buffer containing 0.5 uM oligo-dT primer overlayed with 3 ul Silicon Oil (100 cSt; Sigma-Aldrich). Next, reverse transcription was carried out in 400 nl using 0.75 uM Smart-seq3 TSO. Preamplification of cDNA was performed in 1 ul using SeqAmp polymerase (Takara Bio) for 15 cycles. Subsequently, amplified cDNA was diluted with 9 ul H2O and we used 1 ul for downstream tagmentation using 0.002 ul TDE1 enzyme per cell. Final, indexed libraries were pooled per 384-well plate, cleaned and equimolarly pooled before sequencing.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: mixed pool of 4 individuals
characteristics.characteristic: individual
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: liver
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: C57BL/6N
characteristics.characteristic: strain
characteristics.text: YoungOld_Liver_tissue
characteristics.characteristic: title