GSM5583710: RNA-seq of GFP overexpressing cells rep1; Mus musculus; RNA-Seq
The somatosensory cortex of in utero electroporated E14.5 mouse embryos was isolated and the electroporated area dissected. Single cell suspension was prepared using the papain-based neural dissociation kit (Miltenyi Biotec, Cat. N: 130-092-628) according to the manufacturer protocol with reduced dissociation times (5 min. instead of 15 min. at 37˚C). Cell number as well as the viability was assessed using the Countess™ II Automated Cell Counter (Invitrogen). Dissociated cells were and immediately FAC-sorted at a BD FACSAria Fusion (BD Bioscience) with four lasers (405, 488, 561, 640) using a 100µm nozzle. RNA from sorted cells were isolated using TRIzol (ThermoFisher, Cat. N.: 15596026) according to the manufacturer instructions. RNA-seq libraries were generated using the NEBNext® Single Cell/Low Input RNA Library Prep Kit (New England Biolabs, Cat. N.: E6420) according the manufacturer's instruction.