Illumina MiSeq paired end sequencing
N/A
Diploid cells of the bioethanol isolate Saccharomyces cerevisiae PE-2 (Basso et al. FEMS Yeast Res. 2008 Nov;8(7):1155-63.) were induced to sporulate. A single tetrad was dissected and two spores were isolated. Those correspond to the spore H3 (MATα) and H4 (MATa). Individual cell cultures were prepared from each spore and high quality DNA was extracted (DNeasy Plant Mini Kit, QIAGEN, Germany) from each sample. DNA was enzymatically shredded with the use of NEBNext® dsDNA Fragmentase® (New England Biolabs Inc, USA) and fragments around 500 bp were size selected for library construction (IlluminaTruSeq DNA PCR-Free Library Prep Kit). Libraries derived from H3 and H4 DNAs were sequenced (2x 300 pb) on the Illumina MiSeq platform (Reagent Kit v3, 600-cycles) at the Federal University of Rio Grande do Sul, Brazil).