Schizosaccharomyces pombe Genome sequencing

To explore the intraspecific diversity of the fission yeast Schizosaccharomyces pombe, we performed genome resequencing on 38 S. pombe isolates obtained from four culture collections in China and one culture collection in USA, including 20 strains from CGMCC (China General Microbiological Culture Collection Center), 13 strains from CICC (China Center of Industrial Culture Collection), 1 strain from CICIM (Culture and Information Centre of Industrial Microorganisms of China Universities), 1 strain from CFCC (China Forest Culture Collection Center), and 3 strains from NRRL (United States Department of Agriculture Agricultural Research Service Culture Collection). Single cell-derived clones of these strains were deposited into our laboratory strain collection (DY collection) and given strain names that begin with the initials DY. Cells grown on YES solid medium were used for genomic DNA preparation using the MasterPure Yeast DNA Purification Kit (Epicentre). The kit manufacturer's protocol was followed, with the exception of lysing the cells by glass bead beating in a FastPrep-24 homogenizer (MP Biomedicals) for 20 seconds at a speed setting of 6.4 m/s. The sequencing library for DY15505 was constructed using the NEBNext DNA Library Prep Master Mix (NEB). For the other 37 strains, tagmentation-based sequencing library preparation was performed using home-made Tn5 transposase (Picelli et al., 2014). Post-tagmentation gap filling and PCR amplification were performed using the KAPA HiFi HotStart PCR Kit (Kapa Biosystems) with the following cycling parameters: 3 min at 72°C, 30 sec at 95°C, and then 11 cycles of 10 sec at 95°C, 30 sec at 55°C, and 30 sec at 72°C. AMPure XP beads (Beckman Coulter) were used to select PCR product in the size range of 400 bp to 700 bp. Paired-end sequencing was performed using Illumina HiSeq 2000 (2×96 read pairs), HiSeq 2500 (2×100 read pairs or 2×101 read pairs), or HiSeq X Ten sequencer (2×150 read pairs).