title: RNA-seq of Candida albicans in hyphae inducing conditons treated or not with nicotinamide
design: RNA-seq of Candida albicans in hyphae inducing conditons treated or not with nicotinamide
library_construction_protocol: C. albicans grown in RPMI at 37C and 5% CO2 treated or not with 10 mM NAM was harvested by centrifugation (8,000 g for 10 min at 4°C) and washed with UltraPure™ DNase/Rnase-Free Distilled Water. The cell pellet was resuspended in 1 ml QIAzol Lysis Reagent (Qiagen) and disrupted mechanically with BeadBug microtube homogenizer.  Budding yeast cell culture was diluted at a density of 5x105 cells/mL and seeded in 90-mm diameter Petri dishes  in RPMI 1640 medium with L-Glutamine, and incubated at 37°C, 5% CO2 for 16 h with (treated) or without (CTRL) 10 mM nicotinamide (NAM) Total RNA was purified following QIAzol manufacturer instructions. Three independent biological replicates were performed for either control or treated C. albicans cells. RNA quality was assessed with TapeStation (Agilent) and only RNA with RIN > 8 was used for RNA-seq library production.  Indexed libraries were prepared from 1 µg of purified RNA using TruSeq Stranded Total RNA Sample Prep Kit (Illumina Inc.), according to the manufacturer instructions