RNA-Seq of Homo Sapies: nasal epithelium
Thawed nasal specimens in RNAprotect were agitated to remove cells from the CytoSoft brush either by vortexing or using a TissueLyser without bead (QIAGEN, Hilden, Germany) and then cells were pelleted by centrifugation (5000-10000g, 5 min). Following removal of RNAprotect, the cell pellet was lysed using the QIAzol reagent and total RNA extracted using the miRNeasy Mini Kit (QIAGEN, Hilden, Germany) according to the manufacturers instructions. RNA quantification was performed using the QuantiFluor RNA System (Promega, Madison, WI), and 50 ng of RNA were used as input to the TruSeq RNA Access Library Prep procedure (Illumina, San Diego, CA), which enriches for the coding transcriptome. Libraries meeting quality control criteria for amplification yields were sequenced using NextSeq 500/550 instruments (2x75 bp paired-end reads) with the High Output Kit (Illumina, San Diego, CA).