GSM8084838: mESCs, 50 ng/ml FGF; Mus musculus; RNA-Seq
Cells were washed with PBS and detached with trypsin or Accutase. After centrifugation (5 min at 200xg), cells were resuspended in PBS supplemented with 0.5% BSA. Before GEM generation, cells were passed through a cell strainer (FALCON, 35µm mesh). Droplet generation, lysis, mRNA capture was performed following the instructions by 10x genomics (Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 with Feature Barcoding technology for CRISPR Screening, CG000205 Rev D). Generation of the gene expression libraries was performed following the instructions by 10x genomics (Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 with Feature Barcoding technology for CRISPR Screening, CG000205 Rev D). We chose 12 PCR cycles for cDNA amplification and 13 cycles for the sample index PCR. 10x Genomics single index single-cell RNA sequencing.