GSM5426094: Sample 1_Wild-type zebrafish embryos 24 hours post-fertilisation; Danio rerio; Bisulfite-Seq
24hpf embryos were snap frozen in liquid nitrogen and stored at -80°C prior to DNA extraction. Genomic DNA (gDNA) was extracted from 24hpf embryos using the QIAGEN DNeasy Blood & Tissue Kit (QIAGEN, Chadstone, VIC, Australia) according to manufacturer instructions in two biological replicates. Oxford Nanopore Ligation 1D Kit (Native Barcoding); Zebrafish gDNA was prepared for Nanopore sequencing using the Oxford Nanopore Ligation 1D Kit including 1μL unmethylated lambda phage spike-in (concentration not disclosed by manufacturer) (Oxford Nanopore Technologies, Oxford, UK) according to manufacturer instructions. Briefly, 950ng of zebrafish gDNA was sheared to an average fragment size of 10kb using a Covaris G-Tube (Covaris, Woburn, MA, USA) at 6000rpm for one minute. Long fragment buffer was used to elute DNA fragments following the final AMpure clean-up step. In total, 250ng of the DNA library was loaded onto a PromethION flow cell (pore version 9.4.1, flow cell version FLO-PRO002) and run on a PromethION sequencing device for 64 hours.