characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-05T16:10:11Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-05T16:10:11Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA111438444
characteristics.characteristic: External Id
characteristics.text: NTU
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-05T16:10:11Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-05T16:10:11Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS13531866
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12248:Sample 8
characteristics.characteristic: Submitter Id
characteristics.text: 68
characteristics.unit: year
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: skeletal muscle myoblast
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Commerically available skeletal muscle cells from human donors (old myoblasts were purchased from Promocell (ref.no C-12530) and young myoblasts from Lonza (ref.no CC-2580)) were collected at three time-points across the experiment: at control (CTRL), end of proliferation (END PROL) and end of differentiation (END DIFF) during the repair period; and transferred to low binding/RNase free microcentrifuge tubes containing 350 µL RNeasy lysis Buffer (Qiagen) supplemented with ß-Mercaptoethanol (Sigma-Aldrich) to isolate total RNA using Rneasy Mini Kit (Qiagen). Myoblasts from aged (68 y/o) and young (20y/o) donors were proliferated in growth medium (DMEM+1%pen/strep+20%FBS) for 4 days before switching to differentiation medium (DMEM+1% pen/strep+2%HS+10ug/ml insulin for 6 days to induce myotube formation Differentiated myotubes were incubated with 12% BaCl2 for 2-4h to induce injury. Exposure to chemical injury was followed by the replication of the same experimental protocol used pre-injury. Following lysis, samples were homogenised and loaded onto Rneasy silica membrane for RNA binding. Concentrated and pure RNA was eluted in Rnase-Free water. Concentrations were quantified using spectrophotometry (NanoDrop™ 2000; Thermo Fisher Scientific). RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. To select cDNA fragments of preferentially 370~420 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA). Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system.
characteristics.characteristic: description
characteristics.text: aged post-proliferation
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: DONOR 1
characteristics.characteristic: individual
characteristics.text: in vitro chemical injury
characteristics.characteristic: injury
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: skeletal muscle tissue
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: Sample 8
characteristics.characteristic: title