GSM8208886: WT, day 7; Mustela putorius furo; RNA-Seq
Fully differentiated WT and CF ferret PDE cultures grown at ALI for 2, 5, 7, and 9 days were dissociated using Accutase followed by DNase treatment. Cells were filtered through a 20 uM strainer and pelleted in 0.04% BSA PBS at 500 g for 10 min. Nonviable dead cells were removed by using MACS Dead Cell Removal Kit following 10X Genomics recommendations (Document CG00039). Single cells were counted on a Thermo countess cell counter and 0.04% BSA/PBS was added to achieve a targeted concentration of 1000 cells/ul. Sequencing libraries were generated by following 10X Genomics recommendations (Document CG000315). Briefly, single cells and reverse transcription master mix were partitioned into Gel Beads in partitioning oil in the 10X Chromium controller. After reverse transcription, cDNA libraries were amplified and fragmentated, followed by adaptor ligation and sample index PCR reaction. Libraries were sequenced on NovaSeq 6000 platform by the University of Iowa Genomics Division.