WGS of Strix vara varia: adult female tissue
We used 50 ng genomic DNA to prepare a whole-genome 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 with 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 used a BluePippin (Sage Science) to select library fragments in the size range of 500-700 nt, which, after subtracting the 134 nt of adapters, corresponded to selecting an average insert size of 466 nt. We cleaned the BluePippin products with 0.6X Agencourt AMPure XP (Beckman Coulter) magnetic beads and 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 with 8 cycles PCR. We then cleaned the PCR products with a DNA Clean & Concentrator -5 kit (Zymo Research). We lastly assessed the library fragment size distribution with a 2100 BioAnalyzer (Agilent Technologies) and the concentration of double-stranded DNA material with a Qubit 2.0 Flurometer (Invitrogen). We combined this library with others and sequenced it on two successive runs of 150 nt paired-end sequencing using a 2-lane flow cell on a HiSeq 2500 (Illumina) in rapid mode. On the first run, we obtained sequencing data from a portion of each of the two flow cell lanes. On the second run, we obtained data from a portion of one of the two flow cell lanes. These are the data from lane 1 of run 1.
SUB2419064
CMCB41533_run1_lane1