Sanger Tree of Life HMW DNA Fragmentation: Opentrons® OT-2 for PacBio LI v1

This protocol is for the fragmentation of HMW DNA from either the MagAttract v.3, Plant MagAttract v.5, Automated Plant Organic Extraction, Nanobind tissue or Nanobind nucleated blood Sanger Tree of Life HMW DNA extraction protocols, using the Opentrons® OT-2 liquid handler and a 2 mL 96-well deep-well KingFisher™ plate. This process is highly effective for the fragmentation of DNA extracted from all of the taxonomic groups covered by the Tree of Life Programme, with DNA sheared into an average fragment size range of 12–22 kb for PacBio LI sequencing. The output of this protocol is sheared DNA which can be directed towards the Sanger Tree of Life Fragmented DNA clean up: Automated SPRI protocol v2. This protocol was adapted from settings used for pipette shearing HMW DNA on the Opentrons® Flex as described in ‘Fragmenting High Molecular Weight DNA for PacBio® Long-Read Sequencing Using Pipette Shearing’ by D. Howell et al.