WGS of Heliothis virescens single pupa (PE Illumina)
Genomic DNA (gDNA) from one pupa was submitted to the North Carolina State Genomic Sciences Laboratory (Raleigh, NC, USA) for Illumina paired-end library construction and sequencing. Prior to library preparation, the DNA template was quantified by a Qubit 2.0 Fluorometer (Invitrogen, USA). Library construction was performed using an Illumina TruSeq Nano Library kit according to standard protocol. Briefly, the gDNA was fragmented using a Covaris S2 Ultrasonicator (Covaris, USA) and purified using AMPure XP beads (Beckman Coulter, USA). The fragments were then endrepaired, followed by 550 bp insert size-selection using sequential AMPure XP bead isolation steps. After adapter ligation, the library was enriched by PCR amplification. The amplified library was checked for quality and final concentration using an Agilent 2100 Bioanalyzer (Agilent Technologies, USA) with a High Sensitivity DNA chip before sequencing on an Illumina HiSeq 2500 (100x2 paired end, rapid run). This QC step determined our average insert size was 800bp rather than the target 550bp.