title: WGS of Candida parapsilosis
design: Cells were harvested by centrifuging at 3,000 rpm for 5 min and cell pellets were re-suspended in 200 l Extraction Buffer (2 % Triton X 100, 100 mM NaCl, 10 mM Tris pH 7.4, 1 mM EDTA, 1% SDS). Cells were lysed and nucleic acid was eluted to an aqueous phase by adding ~0.3 g acid-washed beads and 200 l, Phenol/Chloroform/Isoamyl alcohol 25:24:1, agitating with a 600 MiniG bead beater (Spex SamplePrep) for 30 s and centrifuging at 14,000 rpm for 10 min. This aqueous layer was extracted, and the process was repeated twice using 200 l Phenol/Chloroform/Isoamyl alcohol 25:24:1 and 200 l TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA). 80 l 7.5 M Ammonium acetate and 1 ml 100% Isopropanol were added to the aqueous extraction to precipitate the DNA. DNA was pelleted by centrifugation at 14,000 rpm, washed using 70% Ethanol, and air-dried. Pellets were re-suspended in 400 l TE buffer with 1 l RNase A (100 mg/mL) and incubated overnight at 37C. DNA was re-precipitated and washed once more as above and re-suspended in 150 l water.