Sanger Tree of Life HMW DNA Extraction: Manual MagAttract v1
Michelle Strickland, Robin Moll, Clare Cornwell, Michelle Smith, Caroline Howard
Abstract
This protocol describes 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 a wide variety of taxonomic groups covered by the Tree of Life Programme. 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 the genome size of the species, can be directed towards HMW DNA Pooling, HMW DNA Fragmentation: Diagenode Megaruptor® 3 for LI HiFi, HMW DNA Fragmentation: Diagenode Megaruptor® 3 for LI PacBio or HMW DNA Fragmentation: g-Tube for ULI PacBio. Acronyms HMW: high molecular weight SPRI: solid-phase reversible immobilisation HiFi: high fidelity LI: low input ULI: ultra-low input