GSM2576996: Candida_2h_br2 (part 1); Candida albicans; RNA-Seq
Samples of TR146 monolayers for RNA-Seq analysis were collected 2 h and 6 h after infection with Candida. Cells were scrapped into 1 ml chilled PBS and frozen as beads by dropping this suspension into liquid nitrogen. Disruption was carried out using a Mixer Mill MM 200 (RETSCH, Germany) with a shaking frequency of 30/s under cryo conditions. The resulting powder was resuspended in lysis buffer RLTplus (QIAGEN, Germany), supplemented with 0.01% v/v of ß-mercaptoethanol. The extraction of total RNA was performed according to QIAGEN’s Mechanical Disruption Protocol for the isolation of total RNA from yeast, using the RNeasy Plus Mini Kit. The experiments were performed in triplicates. RNA quality and quantity was evaluated using an Agilent 2100 Bioanalyzer with RNA 6000 Nano Chips, following the manufacturer’s protocol. cDNA libraries were prepared using an initial amount of 500 ng of total RNA and Illumina’s TruSeqTM RNA Sample Preparation v2 protocol.