NTHi infection of Normal Human Nasal Epithelia (NHNE) for 14 days
NTHi infection of Normal Human Nasal Epithelia (NHNE) for 14 days Overall design: 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.