GSM8677006: embryoid_EPI_rep6; Mus musculus; RNA-Seq
TRIzol reagent (Invitrogen, 15596026) was added to cell samples to extract RNA. Briefly, 0.5 mL TRIzol reagent was added to wells in a 6-well plate and mixed thoroughly. Next, 0.2 mL chloroform was added (for phase separation), followed by centrifugation at 12,000 g at 4°C for 15 min. The upper aqueous phase was transferred to a new RNase-free tube and an equal volume of isopropanol added. After mixing, RNA was precipitated for 2 h at -20°C, and then centrifuged for 10 min at 12,000 g at 4°C. RNA pellets were washed twice in 75% ethanol and dissolved in RNase-free water. VE and EPI digested by 0.5% pronase E from 5-day embryoids. The SMARTer Stranded Total RNA-Seq Kit v.2 (Takara, 634413) and the VAHTS Universal v.8 RNA-seq Library Prep Kit for Illumina (Vazyme, NR605) were used to prepared total RNA-seq libraries, which were sequenced on an Illumina NovaSeq 6000 platform (Shanghai Jiema).