FRIK1275
Total DNA was prepared using MasterPure Complete DNA and RNA Purification Kit (Epicentre, Madison, WI). RNA was removed by the addition of RNAse A (Thermo Fisher Scientific, Waltham, MA) and  incubation for 30 minutes at 37C. The manufacturers protocol was modified to include a step to precipitate DNA using an overnight incubation in 70% ethanol at -20C. DNA samples were then submitted to the University of Wisconsin-Madison Biotechnology Center. DNA concentration was verified using the Qubit dsDNA HS Assay Kit (Life Technologies, Grand Island, NY). Samples were prepared according to the TruSeq Nano DNA LT Library Prep Kit (Illumina Inc., San Diego, CA) with minor modifications. Samples were sheared using a Covaris M220 Ultrasonicator (Covaris Inc, Woburn, MA), and were size selected for an average insert size of 550 bp using SPRI bead-based size exclusion. The quality and quantity of the finished libraries were assessed using an Agilent High Sensitivity DNA kit and Qubit dsDNA HS Assay Kit, respectively. Libraries were standardized to 2 nM and paired-end 250 bp sequencing was performed using the Illumina MiSeq Sequencer and a MiSeq 500 bp (v2) sequencing cartridge. Images were analysed using the standard Illumina Pipeline, version 1.8.2.