GSM6186011: Sympathoadrenal system E16.5 Wnt1-Cre [SS2_19_344]; Mus musculus; RNA-Seq
Isolated adrenal glands were transferred on a plate with HBSS and cut into small pieces using a clean scalpel. Dissociation was performed using 0,05% Trypsin/0,02% EDTA for 15 minutes at 37°C and triturated with P-1000 and P-200 pipettes tips until complete tissue dissociation. Following the addition of equal volume of 10% FBS, the tissue suspension was centrifugated at 500 g, 5 min, 4°C. Following three washes with 10% FBS the pellet was resuspended in 1xPBS. Cells were sorted using a BD FACSAria III cell sorter into 384-well-plates and stored at -80oC until sequencing took place according to a published protocol (Picelli, Bjorklund et al. 2013). RNA libraries were prepared according to Smart-seq2 protocol.