GSM5960301: xfCM D14 rep 1 [GC1003621]; Homo sapiens; RNA-Seq
Cells from monolayer cultures were released enzymatically from the culture flasks by incubation with TrypLE Express (Life Technologies, UK) for 10 minutes, spun down at 200g for 10 minutes. Microtissues were gently flushed out of the microwells at the indicated time points and spun down at 200g for 10 minutes. Cells and microtissues were lysed in RLT lysis buffer (Qiagen, Germany) + 1% beta-mercaptoethanol (Sigma Aldrich, Germany), vortexed and stored at -80°C. RNA was isolated from the lysates with the RNeasy Mini Kit (Qiagen) according to the manufacturer's protocol. cDNA libraries were generated using the Lexogen QuantSeq FWD Sample Preparation Kit.