GSM5353737: Tag-seq_Human_rep2; Homo sapiens; RNA-Seq
Mitochondria isolation from mouse heart was performed as described previously (PMID: 31588022). For Hela cells, cells at about 80% confluence were harvested, and spin down at 500g for 3min at 4°C, washed once with cold PBS. Cell pellet was resuspended in 1 ml of lysis buffer (20mM Tris-Cl pH7.6, 220mM mannitol, 70mM sucrose, 1mM EDTA, 1xprotease inhibitor, 1mg/ml BSA) and incubated on ice for 15 min, then transferred to glass Dounce homogenizer (1 ml), the cells were broken by 20-25 strokes, followed by centrifugation at 1200g for 3 min at 4°C. The supernatant was transferred to a new Eppendorf tube and spin at 14,000g for 3 min at 4°C to pellet the crude mitochondria. Mitochondrial RNA was prepared from crude mitochondria by Trizol extraction following manufacture's instructions, after ethanol precipitation, the RNA was dissolved in nuclease-free water and treated with RNase-free Dnase I (Thermol) at 37°C for 20 min, followed by purification with Quick RNA Micro prep column (Zymo Research). Library construction was performed by Vertis Biotechnologie (Freising, Germany)