Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.1 v1
Edel Sheerin, Filipa Sampaio, graeme oatley, Maja Todorovic, Michelle Strickland, Raquel Juliana Vionette do Amaral, Caroline Howard
Abstract
This protocol describes the automated extraction of HMW DNA from multiple different tissue samples from a variety of species intended for long-read sequencing, using the Qiagen MagAttract HMW DNA extraction kit and the ThermoFisher KingFisher™ Apex. This process is effective for a wide variety of taxonomic groups covered by the Tree of Life Programme, excluding plants and fungi. The output of this protocol is HMW DNA, which depending upon yield and genome size of the species, can be directed towards either HMW DNA Pooling or HMW DNA Fragmentation: Diagenode Megaruptor®3 for LI HiFi. This protocol was adapted from Sanger Tree of Life HMW DNA Extraction: Manual MagAttract to include automation for a higher throughput of samples, and has since been updated to Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.2 to include a pre-shear SPRI of the HMW DNA extracted. Acronyms HMW: high molecular weight SPRI: solid-phase reversible immobilisation HiFi: high fidelity