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

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 protoc...

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Main Authors: Bein, Bernhard, Chrysostomakis, Ioannis, Arantes, Larissa S., Brown, Tom, Gerheim, Charlotte, Schell, Tilman, Schneider, Clément, Leushkin, Evgeny, Chen, Zeyuan, Sigwart, Julia, Gonzalez, Vanessa, Wong, Nur Leena W.S., Santos, Fabricio R., Blom, Mozes P.K., Mayer, Frieder, Mazzoni, Camila J., Böhne, Astrid, Winkler, Sylke, Greve, Carola, Hiller, Michael
Format: Article
Language:English
Published: BioMed Central 2025
Online Access:http://psasir.upm.edu.my/id/eprint/120192/
http://psasir.upm.edu.my/id/eprint/120192/1/120192.pdf
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author Bein, Bernhard
Chrysostomakis, Ioannis
Arantes, Larissa S.
Brown, Tom
Gerheim, Charlotte
Schell, Tilman
Schneider, Clément
Leushkin, Evgeny
Chen, Zeyuan
Sigwart, Julia
Gonzalez, Vanessa
Wong, Nur Leena W.S.
Santos, Fabricio R.
Blom, Mozes P.K.
Mayer, Frieder
Mazzoni, Camila J.
Böhne, Astrid
Winkler, Sylke
Greve, Carola
Hiller, Michael
author_facet Bein, Bernhard
Chrysostomakis, Ioannis
Arantes, Larissa S.
Brown, Tom
Gerheim, Charlotte
Schell, Tilman
Schneider, Clément
Leushkin, Evgeny
Chen, Zeyuan
Sigwart, Julia
Gonzalez, Vanessa
Wong, Nur Leena W.S.
Santos, Fabricio R.
Blom, Mozes P.K.
Mayer, Frieder
Mazzoni, Camila J.
Böhne, Astrid
Winkler, Sylke
Greve, Carola
Hiller, Michael
author_sort Bein, Bernhard
building UPM Institutional Repository
collection Online Access
description 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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institution Universiti Putra Malaysia
institution_category Local University
language English
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publisher BioMed Central
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spelling upm-1201922025-09-24T08:00:59Z http://psasir.upm.edu.my/id/eprint/120192/ Long-read sequencing and genome assembly of natural history collection samples and challenging specimens Bein, Bernhard Chrysostomakis, Ioannis Arantes, Larissa S. Brown, Tom Gerheim, Charlotte Schell, Tilman Schneider, Clément Leushkin, Evgeny Chen, Zeyuan Sigwart, Julia Gonzalez, Vanessa Wong, Nur Leena W.S. Santos, Fabricio R. Blom, Mozes P.K. Mayer, Frieder Mazzoni, Camila J. Böhne, Astrid Winkler, Sylke Greve, Carola Hiller, Michael 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. BioMed Central 2025 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/120192/1/120192.pdf Bein, Bernhard and Chrysostomakis, Ioannis and Arantes, Larissa S. and Brown, Tom and Gerheim, Charlotte and Schell, Tilman and Schneider, Clément and Leushkin, Evgeny and Chen, Zeyuan and Sigwart, Julia and Gonzalez, Vanessa and Wong, Nur Leena W.S. and Santos, Fabricio R. and Blom, Mozes P.K. and Mayer, Frieder and Mazzoni, Camila J. and Böhne, Astrid and Winkler, Sylke and Greve, Carola and Hiller, Michael (2025) Long-read sequencing and genome assembly of natural history collection samples and challenging specimens. Genome Biology, 26 (1). art. no. 25. pp. 1-25. ISSN 1474-7596; eISSN: 1474-760X https://genomebiology.biomedcentral.com/articles/10.1186/s13059-025-03487-9 10.1186/s13059-025-03487-9
spellingShingle Bein, Bernhard
Chrysostomakis, Ioannis
Arantes, Larissa S.
Brown, Tom
Gerheim, Charlotte
Schell, Tilman
Schneider, Clément
Leushkin, Evgeny
Chen, Zeyuan
Sigwart, Julia
Gonzalez, Vanessa
Wong, Nur Leena W.S.
Santos, Fabricio R.
Blom, Mozes P.K.
Mayer, Frieder
Mazzoni, Camila J.
Böhne, Astrid
Winkler, Sylke
Greve, Carola
Hiller, Michael
Long-read sequencing and genome assembly of natural history collection samples and challenging specimens
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
url http://psasir.upm.edu.my/id/eprint/120192/
http://psasir.upm.edu.my/id/eprint/120192/
http://psasir.upm.edu.my/id/eprint/120192/
http://psasir.upm.edu.my/id/eprint/120192/1/120192.pdf