Generic sample from Saccharomyces cerevisiae
Saccharomyces cerevisiae
Xylose positive segregants of Lalvin AC S. cerevisiae were crossed once to S288c and the resulting diploids were then sporulated.  F2 segregants were scored for xylose utilization in a TECAN plate reader.  1.5mL of overnight YPD culture of each segregant grown was spun down, resuspended, and frozen in 300µL of sorbitol solution (0.9M sorbitol, 0.1M Tris pH 8, 0.1M EDTA).  Samples were pooled by phenotype at this stage and genomic DNA was extracted as described (Treco, D.A., Preparation of Yeast DNA. Curr. Protoc. Mol. Biol. Vol. 5:13.11. 1989).  The pool contained 19 positive segregants.  5µg of genomic DNA were prepared for sequencing using the Illumina Genomic DNA Sample Kit.  Flow cells were prepared using the Illumina Standard Cluster Generation Kit v2, and samples were sequenced on the Illumina Genome Analyzer II.  GAII data were analyzed with the Illumina 1.3.2 pipeline.