Both FRISCR purified mRNA and live single cells were prepared for single-cell transcriptomics using SmartSeq2 (Picelli et al., 2013). After reverse transcription with ProtoScript II (New England Biolabs) and template switching, we amplified cDNA with KAPA HotStart HIFI 2 × ReadyMix (Kapa Biosystems) using 19–22 cycles. We purified PCR products using Ampure XP beads (Beckman Coulter), and quantified cDNA using a High Sensitivity DNA Chip on a Bioanalyzer 2100 (Agilent). We used 1 ng of cDNA to generate RNA-Seq libraries using the Nextera XT library prep system (Illumina) and sequencing of human cortical cells occurred on the Illumina HiSeq using 50 base paired-end reads.
