title: Exp4_#3_wt
organism: Mus musculus
description: Protocols: Pdx1GFP/+; Hes1+/− mice (Ishibashi et al., 1995; Micallef et al., 2005) were time mated with Hes1+/− mice and E10.5 embryos were isolated and head and tail regions discarded. The samples were treated individually with Liberase TL (Roche) and DNAse I (Thermo) in DMEM (Thermo) for 20 minutes at 37°C, with shaking 1100 rpm on a thermomixer to dissociate the cells and subsequently spun down at 500G for 5min. The cells were suspended in 0.125% Trypsin-EDTA (Thermo) and after 10 min at 37°C the trypsinization was stopped with FBS. Then the cells were spun down at 500G for 5min and suspended in FACS Buffer (2% FBS in PBS with DAPI). Each embryo sample was sorted separately on a Sony SH800 cell sorter directly into RLT plus (Qiagen). We retrieved 200-500 GFP-positive cells per embryo. RNA was purified with RNeasy Plus Micro (Qiagen) according to the manufacturer's instruction. RNA quality was verified using a Bioanalyser Pico Kit (Agilent). libraries were generated using the Ovation SoLo RNA-seq system (NuGen) according to the manufacturer's instructions.