CrossRef Open Access 2025

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

Amy Denton Caroline Howard

Abstrak

This protocol is for the fragmentation of HMW DNA from either the MagAttract v.3, Nanobind tissue or Nanobind nucleated blood Sanger Tree of Life HMW DNA extraction protocols, using the Opentrons® OT-2 liquid handler. This process is highly effective for the fragmentation of DNA extracted from bird and fish species covered by the Tree of Life Programme, with DNA sheared into an average fragment size range of 30–70 kb for ONT 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 v2 protocol. 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. and from settings for pipette shearing HMW DNA on the Opentrons® OT-2 as detailed in the Sanger Tree of Life HMW DNA Fragmentation: Opentrons® OT-2 for PacBio LI protocol.

Penulis (2)

A

Amy Denton

C

Caroline Howard

Format Sitasi

Denton, A., Howard, C. (2025). Sanger Tree of Life HMW DNA Fragmentation: Opentrons® OT-2 for ONT v1. https://doi.org/10.17504/protocols.io.eq2ly49omlx9/v1

Akses Cepat

Informasi Jurnal
Tahun Terbit
2025
Bahasa
en
Sumber Database
CrossRef
DOI
10.17504/protocols.io.eq2ly49omlx9/v1
Akses
Open Access ✓