GSM4845006: HH8 NP negative rep2; Gallus gallus; RNA-Seq
HH4 embryos were transfected with neural plate specific enhancers (listed above) and developed to HH8. Embryo heads were dissected and dissociated in Accumax (Accutase, SCR006) cell dissociation solution for 20 minutes at RT under mild agitation. Dissociated cells were FACs sorted directly into 100ul of lysis buffer form the RNAqueous-Micro Total RNA Isolation Kit (Thermo Fisher Scientific. Am1931) (5,000 cells per smaple). Total RNA was isolated according to manufacturer's protocol. RNA was poly(A) selected using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, E7490). TruSeq-barcoded RNA-seq libraries were generated with the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, E7760) and sequences on an Illumina NextSeq 500 instrument with 150bp paried end reads.