UCE target enrichment of Pachystigmus occipitalis
Genomic DNA was extracted from ethanol-preserved and pinned specimens using a non-destructive technique (Ceccarelli et al. 2012) with the EZ-10 Spin Column Genomic DNA minipreps Kit (BIOBasic, Toronto, ON, Canada). For library preparation, up to 150 ng of input DNA per sample was resuspended in 100 l of ultrapure water. DNA was sheared into ~200600 bp fragments using a BioRuptor Pico sonicator. Libraries were prepared following the protocol described by Branstetter et al. (2017), using the Kapa Hyper Prep Kit (Kapa Biosystems Inc., Wilmington, MA, USA) and custom TruSeq-style dual-indexing barcodes adapters (Glenn et al. 2019) for in silico demultiplexing. Samples were pooled at equimolar concentrations in groups of 710 libraries for enrichment, with a total input of 2000 ng of DNA per enrichment reaction. The UCE enrichment was performed using the RNA probe set Hym v.2 designed for Hymenoptera.
SUB15089778
CNIN4688