GSM3039265: WT rep1; Mus musculus; RNA-Seq
E10.5 mouse embryos were harvested in cold PBS and were digested in 0.1% Trypsin for 15 min at 37 °C with intermittent agitation. Cells were resuspended in sorting buffer (PBS, 1% fetal calf serum, 1 mM EDTA), filtered through a 35 μm nylon mesh (Corning), and stained with the viability dye 7AAD (Biolegend). FACS-based isolation of GFP-positive cells was performed on instruments at the Stanford Shared FACS Facility. After sorting, cells were pelleted and lysed in TRIzol (ThermoFisher) and RNA was extracted using the RNeasy mini kit (Qiagen).  RNA quality was assessed using a Bioanalyzer, and only samples with an RNA integrity number (RIN) greater than 8.0 were used for library preparation. RNA-Seq libraries were prepared using the TruSeq RNA Sample Preparation Kit (Illumina). Each library was prepared from 300ng to 1ug of RNA that had been obtained from FACS-isolated cells from a single E10.5 embryo.
GEO Accession
GSM3039265