title: GSM5471648: neurons, N/D/S replicate #2; Mus musculus; RNA-Seq
library_construction_protocol: Day 12 EBs were collected into a 15mL conical tube and spun down at 200xg for 5 minutes. EBs were then suspended in 500uL of 1x AccuMax (Gibco) enzyme solution with 10 Units DNAaseI (Thermo) and incubated at 370C for lysing. Lysate was quenched with 5mL of HEPES buffered saline and triturated into a single cell suspension and filtered with a 70um cell strainer. tdTomato+ differentiated neurons were sorted by flow cytometry (BD Influx System) directly into Trizol. mRNA was purified using an RNA miniprep kit (Zymo) and measured with a Bioanalyzer (Agilent) to ensure an RIN score of 7 or greater. mRNA was enriched using a standard poly-A-pull-down prior to library construction with Illumina TruSeq chemistry. The library is sequenced using Illumina NovaSeq 6000.