GSM8329228: N/TERT-2G cells, hFWE KO Clone #5, replicate 1; Homo sapiens; RNA-Seq
Total RNA was isolated from epidermal organoids after 10 days of air liquid interface culture using the Favorprep total tissue RNA kit (Favorgen Biotech, Tung, Taiwan) according to manufacturer's instructions. To prepare sequencing libraries, a total RNA input of 500 ng was used for library preparation with the NEBNext Ultra II Directional RNA Library Prep kit for Illumina along with NEBNext Poly(A) mRNA Magnetic Isolation Module (both New England BioLabs Inc.) according to the manufacturer's protocol. The library quality and quantity of RNA were assessed using RNA ScreenTape Assay (Agilent) and Agilent 4200 TapeStation System following the manufacturer's protocol. Libraries were pooled equimolar for sequencing to a final concentration of 3 nM. The pooled libraries were sequenced on the Illumina NextSeq 2000 system (Illumina) using NextSeq 1000/2000 P2 Reagent Cartridge 100 cycles and P2 Flow Cell (Illumina).