This is our \u2018one-pot ligation\u2019 protocol for Oxford Nanopore ligation libraries. It benefits from increased recovery of library and faster preparation time by using the Ultra II ligation module\u00a0in conjunction\u00a0with the Ultra II end repair\/dA-tailing module therefore removing a clean-up step. It can be used with or without the optional FFPE DNA repair step which shoudl help to\u00a0improve read-lengths in nicked DNA. The incubation times given\u00a0here are sufficient to generate high quality libraries in under one hour. In the past we have cut these times in\u00a0half without noticably impacting performance but we will leave this up to you.