RNA-seq of human skin: effects of isotretinoin on non-lesional skin in severe acne patients
Total RNA was isolated from full-thickness skin biopsies using the Qiagen RNeasy Fibrous Tissue Mini Kit according to the manufacturers protocol. RNA concentration was measured with a Qubit 3 Fluorometer, and RNA quality was assessed using an Agilent TapeStation system prior to sequencing library preparation. Libraries were prepared using the Illumina Stranded mRNA Kit and sequenced on an Illumina NextSeq 2000 platform, generating approximately 20-30 million 100 bp single-end reads per sample.