WGS of S. cerevisiae GHP1
High quality, pure genomic DNA (8 microgram) was fragmented down to average 20 kb size using the Covaris g-TUBE device  (Cat # 520079), and following the manufacturers recommendations. This step was followed by AMPure clean-up (1:1 bead:sample ratio). Five micrograms of sheared and concentrated DNA (140 ng/ul) were used for the subsequent SMRT bell library construction steps. The details for this procedure are described in PacBio document P/N 100-286-000-07. This procedure resulted in 1.2-1.5 microgram of SMRT bell library (i.e., 25-30% yield). The final library was size-selected in the SageELFTM instrument (Cat# ELD 7510), using 0.75% agarose gel cassettes and the 1-18 kb v2 cassette definition program. Typically, fractions in wells 2 and 3 contained library fragments in the 15-25 kb range. The desired SageELF fractions were cleaned using AMPure magnetic beads (0.6:1.0 beads to sample ratio) and eluted in 15 ul of 10 nM Tris HCl, pH 8.0. Library fragment size was estimated by the Agilent TapeStation (genomic DNA tapes), and these data were used for calculating molar concentrations. Between 100-125 pM of library was loaded onto the PacBio RS II sample plate for sequencing, using P6/C4 chemistry and 4-hr movies. All other steps in for sequencing were done according the recommended protocol by the PacBio sequencing calculator and the RS Remote Online Help system. One SMRT cell run generated 85-95 thousand reads with an average polymerase read length (post filter) of 18 kb and average sequencing output of 3.3 Gb.