Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.2 v1
Maja Todorovic, graeme oatley, Caroline Howard
Abstract
This protocol describes the automated extraction of HMW DNA from cryogenically homogenised tissue samples from plants and fungi intended for long-read sequencing. It employs the Qiagen MagAttract HMW DNA extraction kit and the Thermo Fisher KingFisher™ Apex. This process is effective for a wide variety of plant species covered by the Tree of Life Programme. 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: Automated Plant MagAttract v.1 to improve sample lysis, and was updated to Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.3 to include a pre-shear SPRI of the HMW DNA extracted. Acronyms HMW: high molecular weight SPRI: solid-phase reversible immobilisation HiFi: high fidelity