GSM5460953: Chick brachial motor neurons - RNA-seq 2; Gallus gallus; RNA-Seq
For mouse, spinal cords of Hb9-GFP (Jackson laboratory) were dissociated and GFP+ MNs were manually collected. The regions for forelimb MNs were visualized under a fluorescence dissection scope and separated from other neurons using a sharp scalpel. The spinal cords were digested with pronase for 30min at RT.    For chick MN RNA sequencing, HH stage 12-13 chick embryos were electroporated with a plasmid that drives nuclear YPF expression under the control of Hb9 promoter. After 3day incubation, limb level spinal cords were isolated and dissociated. 5000-23000 YFP+ MNs were collected using a FACS sorter (BD, FACSAria III) and PicoPure RNA Isolation Kit (Arcturus, KIT0202) was used to extract total RNAs. cDNA was synthesized and amplified using Ovation RNA amplification system v2 (Nugen, 3100-12). The sequencing library was prepared using the TruSeq RNA sample prep Kit (Illumina, CA).