GSM2339931: Drd2 GFP#18; Mus musculus; RNA-Seq
Transgenic mice were sacrificed, brains were extracted in ice-cold ACSF with activity blockers (TTX, APV, DNQX), 400 µm brain slices containing the Dorsal Raphe were sectioned on a vibratome, slices were digested in protease for an hour, Dorsal Raphe was microdissected with fine scissors, triturated into a single-cell suspension, cells were plated on a dish, and GFP+ and mCherry+ cells were manually sorted using a glass pipette controlled by mouth aspiration under a dissection microscope with fluorescence optics. Sorted single-neurons were deposited in a 250 µm PCR tube containing 50 µl exraction buffer from the PicoPure RNA Isolation Kit (Arcturus), heated for 30 minutes at 42C and stored at -80C until further processing. Total RNA was purified using the PicoPure RNA Isolation kit (Arcturus), including an on-column DNA digestion step, cDNA was synthesized and amplified using the Ovation RNA-seq V2 kit (Nugen), cDNA was sheared into ~200bp fragments by Covaris sonication, and RNA-seq libraries were prepared using the Ovation Ultralow Library System (Nugen)
GEO Accession
GSM2339931