Illumina HiSeq 2000 paired end sequencing; Mistranslation accelerates the adaptation of Candida albicans to Fluconazole
Mistranslation accelerates the adaptation of Candida albicans to Fluconazole
Resistance to the antifungal drug was monitored using the E-test assay method (AB biodisk, Biomerieux). The experiment was pursued until strains were growing in liquid media containing a Fluconazole concentration of 256ug/ml (suffix FH) DNA extraction was carried out using the Genomic-tip 100/G kit (Qiagen) according to the manufacturer's protocol. DNA quantity and integrity was assessed using the Quant-it Picogreen dsDNA quantitation assay and by agarose gel electrophoresis. Libraries were constructed using Illumina TruSeq DNA Sample Prep standard protocol. Briefly, 5 ug of high molecular weight genomic DNA (gDNA) was fragmented by Covaris sonication device. Following sonication, DNA fragments were end-repaired and A-tailed. Adapters were then ligated via a 3' thymine overhang. Finally, ligated fragments were amplified by PCR (10 cycles). insert sizes were about  400-500 bp as evaluated in an Agilent DNA 1000 Analyzer Chip.