GSM6323468: Airway 2D culture, rep 5; Homo sapiens; RNA-Seq
RNA was extracted using a Zymo Direct-zol RNA MiniPrep Plus Kit. The quantity of the RNA extracts was determined using a DeNovix RNA Fluorescence Assay Kit and DeNovix DS-11 FX+ Spectrophotometer/Fluorometer, and the integrity of each extract was determined using an Agilent 4200 TapeStation and reagents. cDNA libraries were prepared using a Perkin Elmer NEXTFLEX Combo-Seq mRNA/miRNA Kit. Proper sizing of cDNA libraries were validated using an Agilent 4200 TapeStation, and library quantities were determined by qPCR using a KAPA Library Quantification Kit (Roche). The cDNA libraries were then normalized prior to pooling to ensure equal library representation. Pooled cDNA libraries were sequenced using an Illumina NovaSeq 6000 System, generating ~25 million 1 x 100 nt reads.