WGS of Saccharomyces cerevisiae: sgs1 GCR pool 1
Samples for whole genome sequencing were prepared by growing yeast cultures in 5 ml of YPD, followed by DNA extraction using the Yeastar DNA purification kit (Zymogen). Briefly, cells were centrifuged at 500 g, pellet resuspended in YD digestion buffer and R-Zymolyase by vortexing, incubated at 37C until the solution became clear and resuspended in YD lysis buffer by vortexing. Chloroform was added followed by mixing for 2, centrifuged at 14,000 rpm, supernatant loaded onto Zymo-Spin III column, centrifuged and washed thrice with the DNA Wash Buffer. DNA was eluted with 10 mM Tris-Cl pH 8. DNA was quantified by using Qubit 3.0 Fluorometer (Invitrogen, Carlsbad, CA) as well as by NanoDrop 2000c (Thermo Scientific) to check for purity.