NextSeq 500 paired end sequencing; Single cell RNA-seq of mouse E10.5 progenitor and E14.5 barrelette neurons
To collect E14.5 Drg11vPrV/+ post-mitotic barrelette neurons from Drg11tdTomato/+;r2EGFP/+ mice, r2 – r3 derived regions were micro dissected. The boundary between r3 and r4 was identified by the position of the facial nerve. Dissected tissue was kept in PBS 1X containing 2μM  actinomycin D (Sigma, A1410) for 10 minutes on ice, then treated with papain digestion mix (papain 10mg/ml/ cysteine 2.5mM/ HEPES pH7.4 10mM/ EDTA 0.5mM/ DMEM 0.9X/ 40μM actinomycin D) for 3 minutes at 37°C and immediately put on ice.  Tissue was rinsed by ice-cold DMEM 1X containing 2uM actinomycin D, and dissociated by pipetting and filtered. r3-derived cells were collected by FACS. Not Applicable. 8,000 cells were sorted directly into 50µl of PBS-0.04%BSA. All the cells were loaded into one channel of a Chromium Single Cell A Chip (10X genomics). Reverse-transcription, cDNA pre-amplification (13 cycles) and library preparation were performed according to the manufacturer instructions (Chromium Single Cell 3′ Library & Gel Bead Kit v2, 10X genomics). 