DIV-4.2
R1 embryonic stem cells (ESCs) were maintained and differentiated into highly enriched glutamatergic neurons as described in Hubbard, et al (BMC Neuroscience 2012, 13:127).   High quality RNA was isolated from ESCs (DIV -8; n=4); neuroepithelial stem cells (DIV -4; n=3); radial glia (DIV 0; n=3); developmental stage (DS) I/II neurons (DIV 1; n=4); DS III/IV neurons (DIV 7; n=5); and maturing DS IV/V neurons at DIV 16 (n=4); DIV 21 (n=4); and DIV 28 (n=4) using the QIAcube (Qiagen, Valencia, CA) with the RNeasy mini kit protocol.  Reverse-transcribed RNA was prepared for  high throughput sequencing using the TruSeq RNA Prep Kit (Illumina, San Diego, CA).  Paired-end sequence data was generated over 2x50 cycles using an Illumina HiSeq 2000.