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 Preparation Kit (Illumina). 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 KAPA Library Amplification kit (KAPA Biosystems) and then cleaned the reaction with a DNA Clean & Concentrator -5 kit (Zymo Research). We then used a BluePippin (Sage Science) to select library fragments in the size range of 634-709 bp, which, after subtracting the 134 bp of adapters, corresponded to selecting an average insert size of 537.5 bp. We assessed the library fragment size distribution with a 2100 BioAnalyzer (Agilent Technologies). We cleaned the size-selected product with 0.6X Agencourt AMPure XP (Beckman Coulter) magnetic beads to remove adapter dimer of approximately 250 bp in size. We then performed a real-time PCR (rtPCR) using a KAPA Real-Time Library Amplification Kit (KAPA Biosystems) on a CFX96 Touch Real-Time PCR Detection System (Bio-Rad) to amplify the library. We amplified the library with 8 cycles PCR and then cleaned the PCR products with a DNA Clean & Concentrator -5 kit (Zymo Research). We obtained one lane of 300 bp paired-end data sequenced using a MiSeq Reagent Kit v3 on a MiSeq (Illumina). We obtained a second lane of 375 bp read 1 and 225 bp read 2 for a total of 600 bp of paired-end read data sequenced using a MiSeq Reagent Kit v3 on a MiSeq (Illumina).