GSM8524047: WT SC I; Candida albicans; RNA-Seq
Total RNA was isolated from C. albicans cells using phenol-chlororform extraction as previously reported (https://doi.org/10.1128/mbio.03142-21): "For total RNA isolation, cells were resuspended in 440 μL acetate EDTA buffer (50 mM sodium acetate, 10 mM EDTA) with 10% SDS, and 440 μL of acid-phenol-chloroform with isoamyl alcohol (Ambion, Germany) was added. The suspension was thoroughly mixed, incubated for 5 min at 65°C, frozen at −20°C for 30 min, and subsequently thawed at 65°C for 5 min. The phases were separated via 4 min of centrifugation at 16,000 × g, the upper aqueous phase was transferred, and a 1/10 volume of sodium acetate (pH 5.3) and 1 volume of 2-propanol were added to precipitate the RNA for at least 30 min at −20°C. The supernatant was discarded after 10 min of centrifugation at 12,000 × g, and the RNA pellet was washed twice with 80% ethanol and finally solved in 300 μL nuclease-free H2O. The quality and quantity of the RNA were assessed using 2100 bioanalyzer (Agilent Technologies, CA, USA) and Nanodrop (Thermo Fisher Scientific, MA, USA) measurements." Strand-specific cDNA library wer used for sequencing. Libraries were created via poly-A enrichment, mRNA fragmentation, random primed cDNA synthesis, adapter ligation and adapter-specific PCR amplification.