WGS of S. cerevisiae GHP4
Approximately one microgram of genomic DNA (from three yeast isolates) was fragmented down to approximately 500 bp using the Covaris S220 sonicator (Covaris Inc.). DNA (genomic) sequencing libraries were prepared using the NEBNext UltraTMII DNA Library Prep Kit (Cat# E7645S) and the NEBNext Multiplex Oligos  (Cat# E7335S), according to the manufacturers protocol. Library enrichment and barcoding was done by 7 cycles of amplification, and purified by AMPure beads (Beckman Coulter, catalog # A63881).  The final library was quantitated by fluorescence (QUBIT, ThermoFisher), and sized on the Agilent TapeStation. A yield of approximately 150 ng of library was obtained (~650 bp long) with a 400-900 bp fragment size distribution. DNA sequencing libraries were normalized and pooled equimolarly for sequencing on a single Illumina MiSeq, 2x300 cycles run (v3). The library pool was diluted to 4 nM and processed according to the standard Illumina MiSeq sequencing protocol, using 14 pM clustering concentration and 1% PhiX spike-in control. Approximately 50 million high quality (%>=Q30 reads of ~90%) paired-end reads were obtained for a total yield of approximately 15 Gb of sequencing data.