NTHi infection of Normal Human Nasal Epithelia (NHNE) for 14 days

NTHi infection of Normal Human Nasal Epithelia (NHNE) for 14 days Normal human nasal epithelia (NHNE) from five healthy human donors were infected with Haemophilus influenzae strain 86-028NP (MOI 1:100, NHNE:bacteria). Samples for RNA isolation were collected prior to infection (day0) for uninfected NHNE and bacterial pre-cultures, and on day1, 3, 7 and 14 post-infection for NTHi-infected NHNE. RNA was isolated using TRIZOL, treated with EZDnase to remove remaining gDNA and concentrated usingRNA Clean and Concentrate Kits (Zymo). 2-3 microg total RNA (RIN >8.0) were ribodepleted using the RiboMinus kit (Termo Fisher) and the ribodepleted RNA used for library preparation. For all NHNE samples ~ 70 million reads were collected per sample and mapped to human and NTHi reference genomes. for NTHi infected NHNE samples, approximately 2 milion reads could be mapped to the bacterial genome.