GSM3211389: UL_P5_1 [RNA-Seq]; Mus musculus; RNA-Seq
For deep layer neurons, mice expressing the Golli-GFP transgene were used. For upper layer neurons, a birth-dating plasmid expressing Cdk5r promoter-GFP was introduced into the neocortex using in utero electroporation at E15.5, during the peak of upper layer neurogenesis. Neocortex was micro-dissected from the embryonic brain and kept on ice. A total of 2-8 embryos were used per replicate. Cells were dissociated in papain, and GFP-positive cells were purified using FACS. Cells were sorted into micro-centrifuge tubes containing neurobasal/DMEM-based cell culture medium and kept cold. Total RNA was extracted using a Qiagen RNeasy Micro kit. Illumina TruSeq RNA Sample Prep Kit (Cat#RS-122-2101) was used with a minimum of 100 ng of total RNA for the construction of sequencing libraries. RNA libraries were prepared for sequencing using standard Illumina protocols.