title: NextSeq 500 sequencing; RNA-seq of XTE cells and vesicles, XEN and XEN-like cells, and ES cells
design: RNA-seq of XTE cells and vesicles, XEN and XEN-like cells, and ES cells
library_construction_protocol: Cells were collected from culture using standard procedures.  Mouse embryonic stem cells were maintained in the presence of 2i/Lif (0.4 µM PD0325901, 3 µM CHIR99021 and 4 ng/ml Lif), on plastic dishes coated with 0.2% gelatine.  Total RNA was isolated using Macherey-Nagel RNA isolation kit. 1ug of this was used for library preparation and sequencing.  Enrichment of mRNA was performed using NEBNext® Poly(A) mRNA Magnetic Isolation Module and cDNA library was constructed using NEBNext® Ultra™ II Directional RNA Library Prep Kit.