Dec 08, 2020 McGill Nanopore Ligation LibPrep Protocol SQK-LSK109 Sarah J Reiling , Shu-Huang Chen , Ioannis Ragoussis McGill University dx.doi.org/10.17504/protocols.io.bpegmjbw Shu-Huang Chen 1 1 1 1 2 This protocol is used to for both amplicon and high molecular weight DNA library preparations from genomic DNA using Nanopore ligation kit (SQK-LSK109). It contains no fragmentation and no PCR step. However, a fragmentation step is optional. DOI dx.doi.org/10.17504/protocols.io.bpegmjbw Sarah J Reiling, Shu-Huang Chen, Ioannis Ragoussis 2020. McGill Nanopore Ligation LibPrep Protocol SQK-LSK109. protocols.io protocols.io https://dx.doi.org/10.17504/protocols.io.bpegmjbw protocol , Nov 04, 2020 Dec 08, 2020 44200 For ligation library preparations, it is highly recommended to check DNA's length, quantity and purity. Poor DNA reduces subsequent adapter ligation efficiency, increases the prevalence of chimeric reads, and facilitate pore blocking on sequencing. DNA repair and end-prep 35m 1 Prepare 1.5 ug of genomic DNA into a 1.5 ml Eppendorf DNA LoBind tube. Adjust the volume to 48 uL with nuclease-free water. Mix by flicking the tube and spin down briefly. 1.5 ug genomic DNA is the default amount. We are aiming to have 5-50 fmol for MinION / 1 Citation: Citation: Sarah J Reiling, Shu-Huang Chen, Ioannis Ragoussis McGill Nanopore Ligation LibPrep Protocol SQK-LSK109 https://dx.doi.org/10.17504/protocols.io.bpegmjbw This is an open access protocol distributed under the terms of the Creative Commons Attribution License Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited