RNA-Seq of Homo Sapiens: non-CF bronchial epithelial culture
Total mRNA was extracted using RNeasy Plus Micro Kit, following enclosed instructions. After measuring the spectrophotometric quality of extracted RNA through 260/280 ratios of 2.0 and 260/230 ratios of 1.8-2.2, mRNA samples used to reverse transcribe 1 g of cDNA using iScriptTM cDNA Synthesis Kit. Quantitative real-time PCR was performed with PowerUP SYBR Green Mastermix Master Mix on ViiA7 (Applied Biosystems). RNA libraries were generated using NEB Ultra II Directional mRNA and XX,XXX,XXX 125bp paired-end sequences generated from each library on the Illumina HiSeq 2500 platform using high-throughput V4 flowcells
HB9