Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.2 v1
Graeme Oatley, Amy Denton, Caroline Howard
Abstract
This protocol describes the automated extraction and SPRI 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 Thermo Fisher 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, HMW DNA Fragmentation: Diagenode Megaruptor®3 for LI PacBio or HMW DNA Fragmentation: g-Tube for ULI PacBio. This protocol was adapted from Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.1 to include a pre-shear SPRI of the HMW DNA extracted. Acronyms HMW: high molecular weight SPRI: solid-phase reversible immobilisation LI: low input ULI: ultra-low input