Illumina HiSeq 2500 paired end sequencing; RNA-seq analysis of EpiSCs undergoing differentiation induced by EB formation
RNA-seq analysis of EpiSCs undergoing differentiation induced by EB formation
GOF18-EGFP EpiSC (E3) were maintained in a 1:1 mixture of mouse embryonic fibroblast-conditioned media (MEF-CM) containing 20% KnockOut Serum Replacement (KSR) supplemented with 10 ng/ml FGF2 and N2B27 supplemented with 10 ng/ml FGF2 and 10 ng/ml Activin A (CM/N2B27+FA) on fetal bovine serum (FBS)-coated dishes. For EB formation, 3,000 cells were seeded into 96-well V-bottom plates in DMEM containing 10% FBS, 0.1% poly(vinyl alcohol), and 5 μM Y-27632. On days 1‒5, 16 EBs were pooled and total RNA was isolated using the RNeasy Mini Kit (QIAGEN) with on-column DNase treatment. Sequencing libraries were prepared according to the TruSeq RNA Sample Preparation protocol (Illumina) with poly-A selection.
Experimental Factor: 10% FBS (EB)
growth condition
Experimental Factor: 1
time