Pacbio_WGS_of_yeast_strain:2210
Yeast cells were grown in 50 ml of YPDA (starting at OD of 0.1 at 600 nm) until they reached the exponential phase (two doublings), washed and resuspended in 3 ml of 1 M sorbitol. They were then converted to spheroplasts by adding 50 l of a solution of 4 mg/ml Zymolyase 100T (United States Biological) in 50 mM Tris-HCl, pH 7.5, 1 mM EDTA and 50% glycerol, and incubating at 30C with gentle shaking (70 rpm) for 1 hour, followed by a wash with 1 M Sorbitol. High molecular weight DNA was extracted from the pelleted spheroplasts using MagAttract HMW DNA kit (Qiagen) by following the manufacturers instructions. DNA was sequenced by Novogene on the PacBio sequel platform.