Long-read sequencing and genome assembly of natural history collection samples and challenging specimens

Abstract Museum collections harbor millions of samples, largely unutilized for long-read sequencing. Here, we use ethanol-preserved samples containing kilobase-sized DNA to show that amplification-free protocols can yield contiguous genome assemblies. Additionally, using a modified amplification-bas...

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Main Authors: Bernhard Bein, Ioannis Chrysostomakis, Larissa S. Arantes, Tom Brown, Charlotte Gerheim, Tilman Schell, Clément Schneider, Evgeny Leushkin, Zeyuan Chen, Julia Sigwart, Vanessa Gonzalez, Nur Leena W. S. Wong, Fabricio R. Santos, Mozes P. K. Blom, Frieder Mayer, Camila J. Mazzoni, Astrid Böhne, Sylke Winkler, Carola Greve, Michael Hiller
Format: Article
Language:English
Published: BMC 2025-02-01
Series:Genome Biology
Subjects:
Online Access:https://doi.org/10.1186/s13059-025-03487-9
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author Bernhard Bein
Ioannis Chrysostomakis
Larissa S. Arantes
Tom Brown
Charlotte Gerheim
Tilman Schell
Clément Schneider
Evgeny Leushkin
Zeyuan Chen
Julia Sigwart
Vanessa Gonzalez
Nur Leena W. S. Wong
Fabricio R. Santos
Mozes P. K. Blom
Frieder Mayer
Camila J. Mazzoni
Astrid Böhne
Sylke Winkler
Carola Greve
Michael Hiller
author_facet Bernhard Bein
Ioannis Chrysostomakis
Larissa S. Arantes
Tom Brown
Charlotte Gerheim
Tilman Schell
Clément Schneider
Evgeny Leushkin
Zeyuan Chen
Julia Sigwart
Vanessa Gonzalez
Nur Leena W. S. Wong
Fabricio R. Santos
Mozes P. K. Blom
Frieder Mayer
Camila J. Mazzoni
Astrid Böhne
Sylke Winkler
Carola Greve
Michael Hiller
author_sort Bernhard Bein
collection DOAJ
description Abstract Museum collections harbor millions of samples, largely unutilized for long-read sequencing. Here, we use ethanol-preserved samples containing kilobase-sized DNA to show that amplification-free protocols can yield contiguous genome assemblies. Additionally, using a modified amplification-based protocol, employing an alternative polymerase to overcome PCR bias, we assemble the 3.1 Gb maned sloth genome, surpassing the previous 500 Mb protocol size limit. Our protocol also improves assemblies of other difficult-to-sequence molluscs and arthropods, including millimeter-sized organisms. By highlighting collections as valuable sample resources and facilitating genome assembly of tiny and challenging organisms, our study advances efforts to obtain reference genomes of all eukaryotes.
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spelling doaj-art-e879aceac2c848f0bab2f68949557e8e2025-08-20T02:43:16ZengBMCGenome Biology1474-760X2025-02-0126112510.1186/s13059-025-03487-9Long-read sequencing and genome assembly of natural history collection samples and challenging specimensBernhard Bein0Ioannis Chrysostomakis1Larissa S. Arantes2Tom Brown3Charlotte Gerheim4Tilman Schell5Clément Schneider6Evgeny Leushkin7Zeyuan Chen8Julia Sigwart9Vanessa Gonzalez10Nur Leena W. S. Wong11Fabricio R. Santos12Mozes P. K. Blom13Frieder Mayer14Camila J. Mazzoni15Astrid Böhne16Sylke Winkler17Carola Greve18Michael Hiller19LOEWE Centre for Translational Biodiversity GenomicsCenter for Molecular Biodiversity Research, Leibniz Institute for the Analysis of Biodiversity ChangeBerlin Center for Genomics in Biodiversity Research (BeGenDiv)Berlin Center for Genomics in Biodiversity Research (BeGenDiv)LOEWE Centre for Translational Biodiversity GenomicsLOEWE Centre for Translational Biodiversity GenomicsSenckenberg Research InstituteLOEWE Centre for Translational Biodiversity GenomicsSenckenberg Research InstituteLOEWE Centre for Translational Biodiversity GenomicsGlobal Genome Initiative, National Museum of Natural History, Smithsonian InstitutionInternational Institute of Aquaculture and Aquatic Sciences, Universiti Putra MalaysiaLaboratório de Biodiversidade E Evolução Molecular, Departamento de Genética, Universidade Federal de Minas GeraisMuseum Für Naturkunde, Leibniz Institute for Evolution and Biodiversity ScienceMuseum Für Naturkunde, Leibniz Institute for Evolution and Biodiversity ScienceBerlin Center for Genomics in Biodiversity Research (BeGenDiv)Center for Molecular Biodiversity Research, Leibniz Institute for the Analysis of Biodiversity ChangeMax Planck Institute of Molecular Cell Biology and GeneticsLOEWE Centre for Translational Biodiversity GenomicsLOEWE Centre for Translational Biodiversity GenomicsAbstract Museum collections harbor millions of samples, largely unutilized for long-read sequencing. Here, we use ethanol-preserved samples containing kilobase-sized DNA to show that amplification-free protocols can yield contiguous genome assemblies. Additionally, using a modified amplification-based protocol, employing an alternative polymerase to overcome PCR bias, we assemble the 3.1 Gb maned sloth genome, surpassing the previous 500 Mb protocol size limit. Our protocol also improves assemblies of other difficult-to-sequence molluscs and arthropods, including millimeter-sized organisms. By highlighting collections as valuable sample resources and facilitating genome assembly of tiny and challenging organisms, our study advances efforts to obtain reference genomes of all eukaryotes.https://doi.org/10.1186/s13059-025-03487-9Long-read sequencingPCR amplificationGenome assemblyMuseum collections
spellingShingle Bernhard Bein
Ioannis Chrysostomakis
Larissa S. Arantes
Tom Brown
Charlotte Gerheim
Tilman Schell
Clément Schneider
Evgeny Leushkin
Zeyuan Chen
Julia Sigwart
Vanessa Gonzalez
Nur Leena W. S. Wong
Fabricio R. Santos
Mozes P. K. Blom
Frieder Mayer
Camila J. Mazzoni
Astrid Böhne
Sylke Winkler
Carola Greve
Michael Hiller
Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
Genome Biology
Long-read sequencing
PCR amplification
Genome assembly
Museum collections
title Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
title_full Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
title_fullStr Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
title_full_unstemmed Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
title_short Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
title_sort long read sequencing and genome assembly of natural history collection samples and challenging specimens
topic Long-read sequencing
PCR amplification
Genome assembly
Museum collections
url https://doi.org/10.1186/s13059-025-03487-9
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