Dec 18, 2024

Public workspaceSanger Tree of Life Wet Laboratory Protocol Collection V.2

  • 1Tree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
  • Tree of Life at the Wellcome Sanger Institute
Icon indicating open access to content
QR code linking to this content
Collection CitationAmy Denton, Halyna Yatsenko, Jessie Jay, Karen E Houliston, Caroline Howard 2024. Sanger Tree of Life Wet Laboratory Protocol Collection . protocols.io https://dx.doi.org/10.17504/protocols.io.8epv5xxy6g1b/v2Version created by Amy Denton
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: August 24, 2023
Last Modified: December 18, 2024
Collection Integer ID: 114407
Keywords: reference genome, long read sequencing, sample preparation, tissue dissection, sample homogenisation, PowerMasher, cryoPREP, tissue disruption, cryogenic tissue disruption, plant tissue disruption, tissue homogenisation, cryogenic bead beating, HMW DNA extraction, magnetic bead extraction, automated DNA extraction, KingFisher, MagAttract, manual DNA extraction, plant HMW DNA extraction, solid phase reversible immobilisation, Nanobind, nucleated blood extractions, mollusc extraction, organic extraction, next generation sequencing, third generation sequencing, HMW DNA fragmentation, Megaruptor, low input, PacBio, ultra-low input, Covaris, g-TUBE, SPRI, automated SPRI, manual SPRI, AMPure PB beads, DNA clean-up, RNA extraction, manual RNA extraction, TRIzol, RNA-Seq, Iso-Seq, automated RNA extraction, MagMax, mirVana, magnetic beads, HMW DNA pooling
Funders Acknowledgements:
Wellcome Trust
Grant ID: 218328
Wellcome Trust
Grant ID: 206194
Gordon and Betty Moore Foundation
Grant ID: GBMF8897
Abstract
This is the collection of Sanger Tree of Life Wet Laboratory protocols used for the generation of reference level genome assemblies.

This collection currently contains protocols which cover all stages of the wet laboratory processes performed by the Sanger Tree of Life; these processes are sample preparation, sample homogenisation, HMW DNA extraction, HMW DNA fragmentation, fragmented DNA clean-up and RNA extraction.

This collection will be updated as protocols within the Tree of Life Core Laboratory continue to be developed and refined.



Overview of the segmentation of updated work processes and protocols in the Tree of Life Core Laboratory as part of the Tree of Life Programme at the Wellcome Sanger Institute for reference genome generation.


Files
Protocol
Icon representing the file Sanger Tree of Life Sample Preparation: Triage and Dissection
Name
Sanger Tree of Life Sample Preparation: Triage and Dissection
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life Sample Homogenisation: PowerMash
Name
Sanger Tree of Life Sample Homogenisation: PowerMash
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life Sample Homogenisation:  Covaris cryoPREP® Automated Dry Pulverizer
Name
Sanger Tree of Life Sample Homogenisation: Covaris cryoPREP® Automated Dry Pulverizer
Version 2
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life Sample Homogenisation: Cryogenic Bead Beating of Plants with FastPrep-96
Name
Sanger Tree of Life Sample Homogenisation: Cryogenic Bead Beating of Plants with FastPrep-96
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Manual MagAttract
Name
Sanger Tree of Life HMW DNA Extraction: Manual MagAttract
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.1
Name
Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.1
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.2
Name
Sanger Tree of Life HMW DNA Extraction: Automated MagAttract v.2
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated MagAttract for Small Arthropods
Name
Sanger Tree of Life HMW DNA Extraction: Automated MagAttract for Small Arthropods
Version 1
, Tree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Amy DentonTree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Manual Plant MagAttract v.1
Name
Sanger Tree of Life HMW DNA Extraction: Manual Plant MagAttract v.1
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Manual Plant MagAttract v.2/3
Name
Sanger Tree of Life HMW DNA Extraction: Manual Plant MagAttract v.2/3
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Manual Plant MagAttract v.4
Name
Sanger Tree of Life HMW DNA Extraction: Manual Plant MagAttract v.4
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.1
Name
Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.1
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.2
Name
Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.2
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.3
Name
Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.3
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.4
Name
Sanger Tree of Life HMW DNA Extraction: Automated Plant MagAttract v.4
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file  Sanger Tree of Life HMW DNA Extraction: Automated Plant Organic HMW gDNA Extraction (POE)
Name
Sanger Tree of Life HMW DNA Extraction: Automated Plant Organic HMW gDNA Extraction (POE)
Version 1
, Wellcome Sanger Institute - Tree of Life
Benjamin JacksonWellcome Sanger Institute - Tree of Life
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Hypertonic Washing of Plant Tissue Homogenates
Name
Sanger Tree of Life HMW DNA Extraction: Hypertonic Washing of Plant Tissue Homogenates
Version 1
, Tree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Amy DentonTree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Manual Nucleated Blood Nanobind®
Name
Sanger Tree of Life HMW DNA Extraction: Manual Nucleated Blood Nanobind®
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Automated Nucleated Blood Nanobind®
Name
Sanger Tree of Life HMW DNA Extraction: Automated Nucleated Blood Nanobind®
Version 1
, Tree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Amy DentonTree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Manual Mollusc Nanobind®
Name
Sanger Tree of Life HMW DNA Extraction: Manual Mollusc Nanobind®
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Modified Omega Bio-Tek E.Z.N.A.®
Name
Sanger Tree of Life HMW DNA Extraction: Modified Omega Bio-Tek E.Z.N.A.®
Version 1
, Tree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Amy DentonTree of Life, Wellcome Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Fragmentation: Diagenode Megaruptor®3 for PacBio HiFi
Name
Sanger Tree of Life HMW DNA Fragmentation: Diagenode Megaruptor®3 for PacBio HiFi
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Fragmentation: Diagenode Megaruptor®3 for LI PacBio
Name
Sanger Tree of Life HMW DNA Fragmentation: Diagenode Megaruptor®3 for LI PacBio
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Fragmentation: Covaris g-TUBE for ULI PacBio
Name
Sanger Tree of Life HMW DNA Fragmentation: Covaris g-TUBE for ULI PacBio
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life Fragmented DNA clean up: Manual SPRI
Name
Sanger Tree of Life Fragmented DNA clean up: Manual SPRI
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life Fragmented DNA clean up: Automated SPRI
Name
Sanger Tree of Life Fragmented DNA clean up: Automated SPRI
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life RNA Extraction: Manual TRIzol™
Name
Sanger Tree of Life RNA Extraction: Manual TRIzol™
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life RNA Extraction: Automated MagMax™ mirVana
Name
Sanger Tree of Life RNA Extraction: Automated MagMax™ mirVana
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute
Protocol
Icon representing the file Sanger Tree of Life HMW DNA Extraction: Pooling
Name
Sanger Tree of Life HMW DNA Extraction: Pooling
Version 1
, Wellcome Sanger Institute
Tree of Life Genome Note EditorWellcome Sanger Institute