Illumina WGS of Holacanthus passer: adult female gill
The Illumina library was prepared with 250 ng of DNA using the Kapa Hyperplus Library Preparation kit with only one-third of the volume reactions as described in the manufacturers protocol. The total fragmentation volume was 16.66 ul and was incubated at 37 C for 7:45 min. The incubation parameters were previously optimized to target fragments of ~500 bp. Post-ligation purification was done using a 0.8X KAPA Pure bead cleanup. Library amplification was carried out with a total PCR reaction volume of 16.6 ul for 8 PCR thermal cycles. Finally, we did a double size-selection post-amplification cleanup with SPRIselect beads using a 0.56X upper and 0.72X lower selection ratio (Beckman Coulter, Inc).