Illumina MiSeq paired end sequencing; Transcriptional responses to isobutanol in Saccharomyces cerevisiae wild type and gln3 deletion strains
Transcriptional responses to isobutanol in Saccharomyces cerevisiae wild type and gln3 deletion strains
Cells were harvested at 3,000 × g for 5 min at room temperature. Wild-type BY4741 and BY4741 with gln3 deletion strains were pre-cultivated in liquid SC medium at 30°C for 24 h. Each pre-culture was inoculated into fresh liquid SC medium made inhouse with or without 1.3% (v/v) isobutanol to obtain a starting OD600 of 0.1. Cultivation was performed in test tubes with screw caps at 30°C for 12 h with 240 rpm shaking. Total RNA was extracted from each culture with Isogen-LS (Nippon Gene, Tokyo, Japan). RNA integrity was evaluated for quality control with an Agilent Bioanalyzer 2,100 using Agilent RNA 6,000 Nano Kit (Agilent technologies, Santa Clara, CA). cDNA library preparation was performed using KAPA RNA HyperPrep Kit Illumina Platforms (Kapa Biosystems, Wilmington, MA). Prepared cDNA libraries were validated with an Agilent Bioanalyzer 2,100 using an Agilent High Sensitivity DNA Kit (Agilent Technologies).
Experimental Factor: genotype
wild type genotype
Experimental Factor: growth condition
SC medium without isobutanol