title: WGS of drosophila melanogaster: female adult
design: For PacBio sequencing, the female virgin flies (ISO-1 strain, ~180 individuals) were collected, and starved for 1.5 h and flash-frozen in liquid nitrogen. Genomic DNA was extracted from whole flies of female with SDS, quantified and quality controlled with Nanodrop, Qubit and agarose gel electrophoresis. Purified DNA was performed on library preparation for PacBio sequencing following the Procedure & Checklist of 20 kb Template Preparation Using BluePippin Size-Selection System. Briefly, the DNA was sheared by a Covaris g-TUBE device and purified with AMPure PB beads. The fragmented DNA was subject to DNA damage repairing and end repairing, and ligated with adapters. Then purified ligation products were size selected by BluePippin Size Selection system. After annealing and binding of SMRTbell templates and preparation for MagBead loading,librariey was run on the PacBio RS II system, and the run for each sample was composed of two SMART cells.