HH4 embryos were transfected with neural plate or neural crest 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 (GFP+) directly into 1ml of Trizol (Thermo Fisher Scientific, 15596026)  for total RNA isolation (10,000 cells/experiment). RNA was isolated according to manufacturer's protocol. Small RNA-sequencing libraries were generated using NEB Next Small RNA Library Prep Set for Illumina (NEB, E7330S), according to manufacturer's instructions (total RNA input of 100ng/ul for each library). Libraries were size selected on a Pippin Prep instrument for fragments between 105bp and 155bp according to manufacturer's suggestions. The tru-seq barcoded libraries were sequenced on an Illumina NextSeq 500 instrument with paired-end 37 bp reads.
GEO Accession
GSM4520632