GSM9009991: S1D6; Homo sapiens; RNA-Seq
Cells were collected at the indicated time points during chemical reprogramming and resuspended at a concentration of 1 × 10 6  cells per ml in PBS with 0.04% bovine serum albumin. Using the Single Cell 3′ Library and Gel Bead kit V3.1 (10x Genomics, 1000075) along with the Chromium Single Cell B Chip kit (10x Genomics, 1000074), the cell suspension containing 300–600 viable cells per ml was loaded onto the Chromium single-cell controller (10x Genomics) to create single-cell gel beads in emulsion (GEMs) following the manufacturer's instructions. The captured cells were lysed, and the released RNA was barcoded through reverse transcription in individual GEMs, performed on a S1000TM Touch Thermal Cycler (Bio-Rad) at 53 °C for 45 min followed by 85 °C for 5 min and incubation at 4 °C. scRNA-seq libraries were constructed using a Single Cell 3′ Library and Gel Bead kit V3.1 (10x Genomics) and sequenced on an Illumina NovaSeq 6000 sequencer, with a sequencing depth of no less than 1 × 10 5  reads per cell with paired-end 150-bp reads (PE150).