GSM9745323: PGPC14 iPSC, Day 4, Normoxia, 100 RPM, Sample 3; Homo sapiens; RNA-Seq
Samples taken from culture were immediately centrifuged at 500xg for 5 minutes. Following centrifugation, supernatant was aspirated and cells were resuspended in 2.5 mL RNAlater stabilization solution (Thermo-Fisher, Cat# AM7023). These samples were stored at 4oC for 24 hours, then transferred to -80oC storage until ready for RNA processing and sequencing. Samples were thawed at ambient temperature and purified using a QIAGEN RNeasy Mini Kit (Cat# 74104) following the manufacturers protocols. The final samples were adjusted for target concentrations of 50 ng/µL in 15 µL of RNAse free water (Thermo-Fisher, Cat# 10977015) and submitted to the Centre for Health Genomics and Informatics (CHGI) at the University of Calgary for analysis. RNA library preparation was carried out with a NEBNext Ultra II Directional RNA Library Prep Kit using the manufacturer's protocols (Illumina, Cat# E7760L).