Illumina HiSeq 4000 paired end sequencing; Total RNAseq of human healthy control inferior turbinate and glucocorticoid-treated, aspirin-exacerbated respiratory disease (AERD) nasal polyp nasal brushings
Total RNAseq of human healthy control inferior turbinate and glucocorticoid-treated, aspirin-exacerbated respiratory disease (AERD) nasal polyp nasal brushings
Nasal brushing samples from inferior turbinates of healthy and nasal polyps of AERD patients of cohort 1 were collected under anterior rhinoscopy using the Rhino-Pro Curette (Arlington Scientific, Springville, UT, USA) as described previously (Dhariwal J. et al, American Journal of Respiratory and Critical Care Medicine, 2017; Proud D. et al, The Journal of Immunology, 2004). Nasal brushing samples were directly transferred into RNAprotect (Qiagen, Hilden, Germany) and stored at -80 °C until RNA isolation. Total RNA was extracted according to the manufacturer's instructions (Quick-RNA MicroPrep Kit, Zymo Research, Irvine, CA, USA). RNA integrity number (RIN) was determined with the Agilent 2100 BioAnalyzer (RNA 6000 Nano Kit, Agilent). For library preparation, 1 μg of RNA was depleted for cytoplasmatic rRNAs , fragmented, and reverse transcribed with the Elute, Prime, Fragment Mix . A-tailing, adaptor ligation, and library enrichment were performed as described in the High Throughput protocol of the TruSeq RNA Sample Prep Guide (Illumina). RNA libraries were assessed for quality and quantity with the Agilent 2100 BioAnalyzer and the Quant-iT PicoGreen dsDNA Assay Kit (Life Technologies).