WGS of Heliothis virescens single pupa (MP Illumina)
Genomic DNA from a second pupa was used for Mate-pair sequencing. Prior to library preparation, whole genomic DNA was run out on a 0.5% agarose gel at 130v for 2 hours. Fragments 8kb or larger, as compared with Bioline Hyperladder I, were excised from the gel and purified using a Zymoclean large fragment recovery kit (Zymo....). The DNA sample was submitted to the Michigan State University Research and Technology Support Facility (East Lansing, MI, USA) for mate pairlibrary preparation and sequencing. The DNA library was prepared using an Illumina Nextera Mate Pair Sample Preparation Kit according to standard protocol. While the original intention was to create a mate pair library with a mate span of ~5kbp the resultant library ended up with a mate span of 8kbp. The library was validated and quantitated using Qubit dsDNA assay, Caliper LabChipGX and Kapa Library Quantification qPCR for Illumina Libraries. The library was loaded on one lane of an Illumina HiSeq 2500 High Output flow cell (v4) and sequenced in a 2x125bp paired end format using HiSeq SBS reagents. Base calling was done by Illumina Real Time Analysis (RTA) v1.18.64 and the output of RTA was converted to FastQ format with Illumina Bcl2fastq v1.8.4. A summary of the run output is attached.