Sanger Tree of Life HMW DNA Extraction: Manual MagAttract v3 v1

This protocol is for the manual extraction of HMW DNA from multiple different tissue samples from a variety of species, excluding plants and fungi, intended for long-read sequencing using the Qiagen MagAttract HMW DNA extraction kit. This process is effective for the majority of taxonomic groups covered by the Tree of Life Programme, excluding plants and fungi. This protocol is particularly useful for samples with limited tissue availability, as it has consistently yielded more DNA from these smaller samples than the equivalent Automated method. 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: Opentrons® OT-2 for PacBio LI, HMW DNA Fragmentation: Diagenode Megaruptor® 3 for LI PacBio or HMW DNA Fragmentation: Covaris g-Tube for ULI PacBio. This protocol was adapted from Sanger Tree of Life HMW DNA Extraction: Manual MagAttract to include an overnight elution in order to maximise the yield of DNA, as well as an updated pre-shear SPRI to improve recoveries of HMW DNA. Acronyms HMW: high molecular weight SPRI: solid-phase reversible immobilisation LI: low input ULI: ultra-low input