Sanger Tree of Life HMW DNA Fragmentation: Covaris g-TUBE for ULI PacBio v1
graeme oatley, Filipa Sampaio, Lucy Kitchin, Raquel Juliana Vionette do Amaral, Caroline Howard
Abstract
This protocol describes the centrifugation-mediated fragmentation of HMW DNA from samples prepared via any of the Sanger Tree of Life HMW DNA extraction protocols. The protocol produces fragments in the 8–10 kb size range using the Covaris g-Tube. The sheared DNA is suitable for downstream long read sequencing, including PacBio sequencing after ultra-low input (ULI) library preparation. This process is highly effective for DNA extracts across all taxonomic groups in the Tree of Life Programme. The output of this protocol is sheared DNA, which can be directed towards fragmented DNA clean up, using either the Manual or Automated SPRI protocols. Acronyms and abbreviations HMW: high molecular weight SPRI: solid-phase reversible immobilisation ULI: ultra-low input