Hyb-Seq of Schefflera polychaeta Harms: Reconstructing the complex evolutionary history of the Papuasian Schefflera radiation through herbariomics
Genomic DNA extracted using modified CTAB protocol. Extractions cleaned using AMPure XP beads and sonicated if above 500 bp. Genomic libraries prepared using NEBNext Ultra II with Dual Index Primers. Libraries size-selected to 350 bp except where DNA template highly degraded (< 300 bp). Libraries amplified at 13 to 14 cycles using KAPA HiFi. Libraries multiplexed 11-12 per pool, each containing 500-1000 ng DNA. Pools enriched using Angiosperms-353 myBaits target capture kit. Pools hybridised at 65C for 24 h, then amplified at 14 to 18 cycles. Libraries sequenced in two pools loaded at 16 pM on Illumina MiSeq using v2 (300-cycles) reagent kits.