5-hmc Enrichment 2
DNA from parthenogenetic D. pulex adults was used. 100 ng of extracted DNA was fragmented using MuSeek Library Preparation Kit, Illumina compatible (Thermo Scientific) and purified using GeneJET NGS Cleanup Kit (Thermo Scientific). Libraries were end repaired, purified and 5-hmC was enriched using 5-hmC Enrichment Kit (Thermo Scientific) according to manufacturer’s protocol. Libraries were PCR amplified for 14 cycles and gel size selected for next generation sequencing. The libraries were analyzed using Agilent Bioanalyzer (Agilent) and KAPA library quantification kit (KAPA biosystems). Sequencing was performed using Illumina MiSeq platform with MiSeq reagent kit v3 150-cycle (Illumina).