Mar 25, 2020

Public workspaceProtocols for "Sequencing smart: De novo sequencing and assembly approaches for a non-model mammal"

  • Graham J Etherington1,
  • Darren Heavens1,
  • David Baker1,
  • Ashleigh Lister1,
  • Rose McNelly1,
  • Gonzalo Garcia1,
  • Bernardo Clavijo1,
  • Iain Macaulay1,
  • Wilfried Haerty1,
  • Federica Di Palma1
  • 1The Earlham Institute, Norwich Research Park, Norwich, NR4 7UZ, United Kingdom
  • GigaScience Press
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Collection CitationGraham J Etherington, Darren Heavens, David Baker, Ashleigh Lister, Rose McNelly, Gonzalo Garcia, Bernardo Clavijo, Iain Macaulay, Wilfried Haerty, Federica Di Palma 2020. Protocols for "Sequencing smart: De novo sequencing and assembly approaches for a non-model mammal". protocols.io https://dx.doi.org/10.17504/protocols.io.bd3ri8m6
Manuscript citation:
Graham J Etherington, Darren Heavens, David Baker, Ashleigh Lister, Rose McNelly, Gonzalo Garcia, Bernardo Clavijo, Iain Macaulay, Wilfried Haerty, Federica Di Palma, Sequencing smart: De novo sequencing and assembly approaches for a non-model mammal, GigaScience, Volume 9, Issue 5, May 2020, giaa045, https://doi.org/10.1093/gigascience/giaa045
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 protocol and it's working
Created: March 22, 2020
Last Modified: March 26, 2020
Collection Integer ID: 34641
Keywords: polecat, vertebrate, non-model organism, Illumina, chromium, Bionano, assembly, sequencing,
Abstract
Whilst much sequencing effort has focused on key mammalian model organisms such as mouse and human, little is known about the correlation between genome sequencing techniques for non-model mammals and genome assembly quality. This is especially relevant to non-model mammals, where the samples to be sequenced are often degraded and low quality. A key aspect when planning a genome project is the choice of sequencing data to generate. This decision is driven by several factors, including the biological questions being asked, the quality of DNA available, and the availability of funds. Cutting-edge sequencing technologies now make it possible to achieve highly contiguous, chromosome-level genome assemblies, but relies on good quality high-molecular-weight DNA. Here we use a range of different genomic technologies generated from a roadkill European Polecat (Mustela putorius) to assess various assembly techniques on this low-quality sample. We evaluated different approaches for de novo assemblies and discuss their value in relation to biological analyses. The high degree of variability between each de novo assembly method (assessed from the seven key metrics) highlights the importance of carefully devising the sequencing strategy to be able to carry out the desired analysis. Adding more data to genome assemblies does not always results in better assemblies so it is important to understand the nuances of genomic data integration explained here, in order to obtain cost-effective value-for-money when sequencing genomes.
Files
Protocol
Icon representing the file Long Mate Pair Library Construction Protocol
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Amplification Free Paired End Library Construction Protocol
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Icon representing the file 10x Genomics Library Construction
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Icon representing the file Bionano genome mapping from animal tissue
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Version 1
, The Earlham Institute
Graham EtheringtonThe Earlham Institute