characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-05-24T00:14:45Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-05-24T00:14:45Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112697202
characteristics.characteristic: External Id
characteristics.text: Experimental Research Laboratory Heart and Vascular Center, Semmelweis University
characteristics.characteristic: INSDC center name
characteristics.text: 2023-05-24T00:14:45Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-05-24T00:14:45Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14694777
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12671:Sample 16
characteristics.characteristic: Submitter Id
characteristics.text: 5 to 6
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: Norway rat
characteristics.characteristic: common name
characteristics.text: Protocols: The animals were euthanized by exsanguination. Than, to remove the residual blood cells from myocardial tissue, in situ perfusion with cold physiological saline was applied. After perfusion, the hearts and the lungs of the animals were removed from the thorax and their weights were quickly measured on a scale. This was followed by conservation of left ventricular myocardial tissue. Accordingly, the apex of the left ventricle was cut into small pieces (40-50mg) and subsequently snap frozen in liquid nitrogen. RNA isolation was performed from 40-50mg of left ventricular myocardial tissue using the RNeasy Plus Universal kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions paying special attention to appendix D in order to ensure purification of total RNA including small RNAs. Isolated RNA was subsequently eluted in 30 µl RNase-free water. RNA quality and concentration were determined and reviewed using a NanoDrop® spectrophotometer (Thermo Scientific™, Waltham, MA, USA) to measure the concentration and optical density (OD) 260/280 values of the samples and an Agilent RNA TapeStation® (Agilent Technologies, Santa Clara, USA) to check RNA quality via RINe number. RINe >5 were accepted as input for library preparation. Quantification was performed via absorbance measurements using Qubit RNA broad range (BR) Assay Kit (Thermo Scientific™, Waltham, MA, USA). Library preparation was done using the QIAseq® miRNA Library Kit (Qiagen, Hilden, Germany) following the manufacturer’s instructions. A total of 500ng of RNA per sample was converted into miRNA next generation sequencing (NGS) libraries. Primers containing UMIs were used during reverse transcription. The resulting cDNA was amplified using PCR (13 cycles) with universal forward and sample index assigning reverse primers. The PCR product was then purified. Quality control of the library preparation was done by capillary electrophoresis using the High Sensitivity Tape D1000 (Agilent Technologies, Santa Clara, USA).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: hypertophy
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: 1034-016,95
characteristics.characteristic: individual
characteristics.text: Rattus norvegicus
characteristics.characteristic: organism
characteristics.text: Rattus norvegicus
characteristics.characteristic: organism
characteristics.text: heart left ventricle
characteristics.characteristic: organism part
characteristics.text: Rattus norvegicus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: Sprague-Dawley
characteristics.characteristic: strain
characteristics.text: Sample 16
characteristics.characteristic: title