Starmerella batistae SB001 genome sequencing
For PacBio sequencing, the DNA was extracted from samples using the cetyl trimethyl ammonium bromide (CTAB) method (Wu et al., 2001). For the CTAB method, 1mL of overnight culture was added to 50mg of acid washed glass beads 425600m (Sigma). 1mL of CTAB extraction buffer was then added, and after incubation at 65C. RNAse A treatment was conducted by adding 2L of RNAse A 100g/mL (Qiagen, Germany) and incubating the samples at 37C for 15min. After purification with phenol:chloroform:isoamyl alcohol (25:24:1) the DNA was eluted in 50L of ultrapure distilled water (Invitrogen) and stored at 4C. The DNA quality was assessed using the NanoDrop LiTE Spectrophotometer (Thermofisher Scientific) to be within the quality specification range required by the PacBio protocol. The genomic DNA was adjusted to 10ng/L in 150L and sheared to ~10 kilobase fragments using g-TUBES (Covaris) following the manufacturers instructions. The quality and size of DNA fragments were verified using Fragment Analyzer (Advanced Analytical Technologies) following the DNF-90 protocol. Samples were prepared for sequencing following the Express Template Prep Kit 2.0 protocol (Pacific Biosciences, USA), with multiplexing using the Barcoded Overhang Adapter kit 8A (Pacific Biosciences, USA). DNA libraries were sequenced using the SMRT Cell 1M chips on the Pacific Biosciences Sequel system with 10h data acquisition time.