Nanodrop 2100c

Total RNA from aliquots was extracted using the RNeasy Plant Mini kit (Qiagen) according to manufacturer’s instructions. RNA concentration was determined in a Nanodrop 2100c (Thermo Fisher Scientific, Waltham, MA, USA) and the integrity was evaluated in a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) using the Agilent RNA 6000 Nano kit (Agilent Technologies, Santa Clara, CA, USA). Total RNA from two biological replicates per condition (SEB or fructose) from both fungi was used as input for the TruSeq RNA Sample Prep v2 kit (Illumina, San Diego, CA, USA) and the libraries were sequenced at the USC Epigenome Center (CA, USA) using the Illumina® HiSeq2000 system, producing single-end reads of 50 bp.

Total RNA was isolated from all samples using MiniBEST Plant RNA Extraction Kit (TaKaRa, Dalian, China). Concentration and purity of total RNA were assessed by a micro-spectrophotometer (NanoDrop 2100C, Thermo, Wilmington, DE, USA). The absorbance ratio of samples was greater than 1.8 both at OD₂₆₀/₂₈₀ and OD₂₆₀/₂₃₀. 28S/18S ratio between 1.8 and 2.0 was used for subsequent experiments. 1 μg aliquot of total RNA, treated with DNase I (Invitrogen, Carlsbad, CA, USA), was reverse-transcribed using PrimeScript RT reagent Kit (TaKaRa, Dalian, China) according to operating manual. All cDNAs were diluted 1:10 with ddH₂O for further analyses.