GSM8208869: WT, donor1; Mustela putorius furo; RNA-Seq
Fully differentiated WT and CF ferret PDE cultures grown at ALI for 14 days were dissociated using Accutase followed by DNase treatment. Qiagen RNEasy mini kit was used to extract RNA from cells. Two transwells per donor were combined. RNA samples with RIN>9 were used for library prep and sequencing. 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.