title: GSM8026662: D13Q_LAE_614_48; Homo sapiens; RNA-Seq
library_construction_protocol: The cell dissociation procedure followed the manufacturer's protocols for the 10x Genomics Chromium system with modifications. A 5 mM EDTA in Accutase solution (Sigma) was administered to both apical and basolateral HBE surfaces and cultures incubated for 20 min at 37 ˚C. The cells detached from the Transwell membrane were centrifuged (300g x 2 min at 4 C) and washed twice with 10% fetal bovine serum (FBS, Sigma) in Dulbecco's Modified Eagle Medium (DMEM, Sigma). The cells were then resuspended in DMEM with 10% FBS with gentle pipetting using wide-bore pipette tips and passed through a 40 µm cell strainer (Fisher Science #22363547) to remove clumps. Cell counting and viability were determined using an automated cell counter (Countess 3FL, Thermo). Single-cell suspensions targeting approximately 10,000 cells were loaded onto a 10x Genomics Chromium Next GEM Chip G (10x Genomics) device following the manufacturer's instructions. After GEM generation, the library preparation was performed following the instructions for the 10x Chromium Next GEM Single Cell 3' GEM Library kit v3.1 (10x Genomics).