WGS of Strix occidentalis caurina: adult female blood
We isolated DNA using a Gentra Puregene Kit (Qiagen) and used 50 ng to prepare a genomic library using a Nextera DNA Sample Prep Kit (Illumina-compatible) (Epicentre). After tagmentation, we cleaned the reaction with a DNA Clean & Concentrator -5 kit (Zymo Research). We amplified the reaction for 5 cycles of PCR using a Nextera DNA Sample Prep Kit (Illumina-compatible) (Epicentre) and the Nextera PCR Enzyme (Epicentre). We then cleaned the reaction with a DNA Clean & Concentrator -5 kit (Zymo Research). We used a LabChip XT DNA 750 Assay Kit on a LabChip XT (PerkinElmer) automated nucleic acid fractionation system to select library fragments in the size range of 375-600 bp, which, after subtracting the 141 bp of adapters, corresponds to an average fragment size of 346.5 bp. We performed a final PCR using 5 µL Klentaq LA 10X Buffer with MgCl (Sigma-Aldrich), 1 µL 12.5 µM dNTPs, 1 µL each of two Illumina-adapter-compatible primers at 10 µM, 1 µL KlenTaq LA DNA Polymerase Mix (Sigma-Aldrich), 5 µL library off of LabChip, and water to make a 50 µL reaction volume. We ran the PCR at 94°C for 2 min; then 5 cycles of denaturation at 94°C for 30 s, annealing at 58°C for 30 s, and extension at 72°C for 3 min; and we performed a final extension at 72°C for 5 min. We cleaned the products using a DNA Clean & Concentrator -5 kit (Zymo Research). We sequenced the library using a TruSeq PE Cluster Kit v3-cBot-HS kit and a TruSeq SBS v3-HS kit on a HiSeq 2000 (Illumina) and obtained one lane of 100 bp paired-end data.