GSM7068345
Monolayer cortical differentiation cultures were dissociated with accutase and papain to obtain a single cell suspension. Dissociated neurons were then washed, filtered and resuspended in 0.04% BSA/PBS + Ribolock RNase inhibitor Libraries were constructed with the Chromium Next GEM Single Cell 3' Kit v3.1 (10x Genomics) according to the manufacturer's protocol. In brief, single cell suspensions were loaded onto a Chromium Next GEM Chip G for GEM generation, cDNA synthesis, cDNA amplification, and library preparation of 7,000-15,000 cells. cDNA amplification included 11-12 cycles and 30-240 ng of the material were used to prepare sequencing libraries with 10-16 cycles of PCR. Indexed libraries were pooled equimolar and sequenced on a NovaSeq 6000 in a PE28/88 run using the NovaSeq 6000 S1 or S4 Reagent Kit (100 or 200 cycles) (Illumina).