GFP-lenti-D_S8_L003
The sorted single cells were submitted to CPRIT UTHealth Cancer Genomics Core at The University of Texas Health Science Center at Houston. The single cell capture and library construction were performed by following the 10x Genomics Chromium Single Cell 3 Reagent Kits v3.1 protocol (CG000204). Briefly, cells were resuspended in PBS with 0.04% BSA at a concentration of 700-1200 cells/l and more than 85% viability. The single cells were loaded onto Chromium Next GEM Chip G (PN-1000120) with partitioning oil and barcoded singe cell gel beads. The barcoded and full-length cDNA is produced after incubation of the gel beads-in-emulsion (GEMs) and amplified via PCR for library construction. The library preparation is performed by following the protocol of Chromium Single Cell 3 GEM, Library & Gel Bead Kit v3 (PN-1000121). The quality of the final libraries was examined using Agilent High Sensitive DNA Kit (#5067-4626) on an Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, USA), and the library concentrations were determined by qPCR using Collibri Library Quantification kit (#A38524500, Thermo Fisher Scientific) on a QuantStudio3 (ThermoFisher Scientific). The libraries were pooled evenly and then underwent paired-end sequencing on Illumina NextSeq 550 System (Illumina, Inc., USA) using High Output Kit v2.5 (#20024907, Illumina, Inc., USA).