RNA-seq of human 3D organotypic epithelial raft culture: cervix 1
RNA was isolated from the epidermal compartment (containing only basal and suprabasal keratinocytes) of organotypic raft cultures using Qiagens RNeasy Mini Kit (Catalog No. 74106) after separating them with sterile forceps from their underlying dermal equivalents and flash-freezing with liquid nitrogen (PMID: 33232584). RNA was eluted in nuclease-free water, carry-over DNA was removed with the TURBO DNA-free Kit (Thermo Fisher Scientific, Catalog No. AM1907), then shipped on dry ice to Novogene Corporation (Sacramento, CA). High-quality eukaryotic mRNA libraries were prepared and sequenced on an Illumina platform, yielding >20 million paired-end reads (2x150 bp length) per sample.