Acanthocephala declivis CMF0069
Ultraconserved element target capture of Acanthocephala declivis
Libraries were constructed from genomic DNA (sheared to 200 to 1000 bp) using a KAPA Hyper Prep Kit with half volume reactions, iTru universal adapter stubs, and iTru 8 bp dual-indexes (Glenn et al., 2016). Hydrophobic Sera-Mag SpeedBeads Carboxyl Magnetic Beads were used for all cleanup steps. Cleaned, amplified libraries were quantified with Qubit, subsequently combined in equimolar amounts into 1000 ng pools. A custom MYbaits kit containing a subset of Hemiptera UCE probes designed from pentatomomorphan species (Faircloth, 2017) was used for target enrichment. The hybridization mixture used half volume baits (2.75 uL) with 2.75 uL molecular-grade water. Probes were hybridized with library pools at 65C for 16 to 24 hours. Bait-target hybrids were bound to Dynabeads M-280 Streptavidin beads, washed, and resuspended in 30 uL IDTE. We used 2.5 uL each of 5 uM iTru P5/P7 primers (Glenn et al., 2016) for the post-capture PCR amplification mix. 14 to 17 cycles of post-capture amplification were performed, followed by cleaning. Enriched library pools were pooled in equimolar amounts and sequenced using a single Illumina HiSeq3000 lane (2x100 run) at the University of Floridas Interdisciplinary Center for Biotechnology Research (ICBR).
Acanthocephala declivis CMF0069
SUB7027241
Acanthocephala declivis CMF0069