GSM7312231: D14_EHMT1_gRNA02 [scRNA-seq]; Homo sapiens; RNA-Seq
Cells on Day 14 of differentiation were harvested, washed with FACS buffer (PBS + 2% FBS), and stained with DAPI for 10 minutes at room temperature. Viable DAPI- cells were isolated by FACS using a Sony MA900. scRNA-Seq targeting 5,000 cells per gRNA was performed using the Chromium NextGEM Single Cell 3ʹ Reagent Kit v3.1 (10X Genomics) per the manufacturer's instructions. scRNA-Seq libraries were barcoded with dual Illumina indices. The concentration of each final library was quantified using a High Sensitivity DNA Bioanalyzer Assay (Agilent). scRNA-Seq libraries were equimolar pooled and shallow sequenced on an Illumina MiniSeq to determine the number of high-confidence cell barcodes per library. The scRNA-Seq libraries were then renormalized and deep sequenced on a NextSeq 550 in a 28-10-10-44 read configuration.