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Sanger Tree of Life Fragmented DNA clean up: Manual SPRI v1

Michelle Strickland, Clare Cornwell, Caroline Howard

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Abstract

This protocol describes the manual clean up of fragmented DNA following the Sanger Tree of Life HMW DNA Fragmentation protocols, using PacBio AMPure PB beads. This process is highly effective for the cleaning and removal of shorter fragments from sheared DNA from all of the taxonomic groups covered by the Tree of Life Programme. The output of this protocol is DNA which can be submitted for long read sequencing, including PacBio sequencing following Low Input (LI) or Ultra-Low Input (ULI) library preparation. Acronyms HMW: high molecular weight SPRI: solid-phase reversible immobilisation LI: low input ULI: ultra-low input

Topics & Concepts

Sanger sequencingProtocol (science)Fragmentation (computing)DNAProcess (computing)Tree (set theory)DNA sequencingDNA fragmentationComputer scienceComputational biologyBiologyMedicineGeneticsMathematicsOperating systemAlternative medicinePathologyApoptosisMathematical analysisProgrammed cell deathGenomics and Phylogenetic StudiesMolecular Biology Techniques and ApplicationsEnvironmental DNA in Biodiversity Studies