GSM6592556: COL_24mo_B; Mus musculus; RNA-Seq
Cell Pellets were lysed in 1mL Trizol (Thermo Fisher, # 15596026) and 100 µL of bromo-chlorophenol (Fisher, # AC173530050) were added. Phase separation was done by centrifugation at 12,000 g for 15 min at 4°C. Supernatant containing RNA was collected, RNA precipitated with an equal volume of isopropanol, and washed with 1 ml 75% ethanol. RNA pellet was collected by centrifuging at 7500 g for 5 min and dissolving in 30 ul nuclease free water. RNA concentration was determined with Qubit RNA Broad range kit (Thermo Fisher, # Q10210) and RNA integrity score (RIN) was determined using Agilent RNA 6000 nano kit (Agilent, # 56067-1511). RNA was sent to BGI, who performed library preparation and sequencing. BGI