title: GSM7110890: Differentiated HSPCs day 9 post-electroporation, donor 2, edited with c220_2T>C guide 1, replicate C, bulk RNA-seq; Homo sapiens; RNA-Seq
library_construction_protocol: Following final FACS enrichment to remove debris and purify transduced cells, 5-mL PBS-BSA coated tubes described above were spun down at 400 x g for 5 minutes at 4ºC. The entire supernatant was carefully removed and 1mL of PBS_0.05% BSA was added. Cells were then counted twice using an automated Countess 2 cell counter (Thermo Fisher, I-CACC2). Cell viability as determined by Trypan Blue was always >95%. Cells were again washed with 5mL PBS_0.05% BSA and resuspended at a final concentration of 1,000 cells / µL and kept on ice. Single-cells were immediately processed using v3.1 Chemistry Dual Index kits according to manufacturer's instructions (10X Genomics), using 20µL of cell suspension and 23.2 µL of water on the cell suspension loading step. Libraries were sequenced using a 28 base pair (Read 1), 10 base pair (Index 1), 10 base pair (Index 2), 90 base pair (Read 2) configuration on Nextseq 500, Nextseq 2000 or Novaseq S2/S4 instruments.