title: Genome sequence of Candida vulturna CBS14366
design: Candida vulturna CBS14366T was cultured in 10 ml yeast peptone glucose broth for 3 days at 25C on a rotary shaker (125 rpm). Biomass was collected by centrifugation at 4,000 rpm for 10 min (Centrifuge 5810R; Eppendorf), supernatant was decanted, 1.5 ml CTAB-buffer (see (Gerrits van den Ende & de Hoog, 1999)) containing 1 mg proteinase K (V3021, Promega) was added followed by 60 min incubation at 60C with periodically mixing by vortexing. The sample was cooled to room temperature and 2 ml chloroform:iso-amylalcohol (24:1) was added and subsequently mixed by flipping for 5-10 min. After centrifugation at 4,000 rpm for 10 min the supernatant (~1200 l) was collected in a 2.0 ml DNA low-binding tube (0030108078; Eppendorf). DNA was precipitated by adding 660 l ice-cold 2-propanol (I9516; Sigma-Aldrich) and mixed by flipping the tube. DNA yield was increased by overnight incubation at -20C. After centrifugation at 14,000 rpm (Centrifuge 5430; Eppendorf) for 10 min, supernatant was removed and the pellet was washed with 2 ml 70% ice-cold ethanol. The dried pellet was resuspended in 150 l IDTE-buffer (10 mM Tris, 0.1 mM EDTA, pH 8; IDT), more IDTE was added when the pellet did not dissolve completely. RNA was removed by adding 1 l RNAse Cocktail Enzyme mix (AM2286; ThermoFisher) per 100 l sample and incubated for 1h at 37C. DNA samples were again washed with chloroform:iso-amylalcohol and precipitated with 2-propanol as described above. DNA quality and quantity was measured in triplicate using Qubit and Nanodrop (both ThermoFisher), purity and integrity was checked on 0.8% agarose gel. DNA was stored at -20C until further use. Library preparation was done by the ligation sequencing kit (SQK-LSK108; ONT) followed by the nanopore sequencing run on a MinION flow cell (FLO-MIN106; ONT) according to the manufacturers instructions.