GSM9344883: ILF3_KO_gene_10x_lane2; Homo sapiens; RNA-Seq
Cells were prepared for single-cell RNA-sequencing by resuspension in 1X PBS with 0.04% BSA as detailed in the 10x Genomics Single Cell Protocols Cell Preparation Guide (10x Genomics, CG00053 Rev C). For the WT experiment, cells were then separated into droplet emulsions using the Chromium Controller (10x Genomics) with Chromium Single-Cell 3′ Gel Beads v3 (10x Genomics, PN-1000075) across 3 “lanes”/”GEM groups” following the 10x Genomics Chromium Single Cell 3ʹ Reagent Kits v3 User Guide with Feature Barcode technology for CRISPR Screening (CG000184 Rev C) with the goal of recovering ∼20,000 cells per GEM group.  For the ILF3 KO cells were cells were separated into droplet emulsions using the Chromium Controller (10x Genomics) with Chromium Single Cell 3' HT Gel Bead Kit v3.1 (10x Genomics, PN-1000370) across 2 “lanes”/”GEM groups” following the Chromium Next GEM Single Cell 3' HT Reagent Kits v3.1 User Guide with Feature Barcode technology for CRISPR Screening (CG000421Rev D) with the goal of recovering ∼30,000 cells per GEM group before filtering. For preparation of gene expression and sgRNA libraries, samples were processed according to 10x Genomics Chromium Single Cell 3ʹ Reagent Kits v3 or HT v3.1 User Guide with Feature Barcode technology for CRISPR Screening (CG000184 Rev C, CG000421Rev D). For sequencing, mRNA and sgRNA libraries were pooled to avoid index collisions at a 10:1 ratio.