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<p>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&#xa0;Gb maned sloth genome, surpassing the previous 500&#xa0;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.</p>

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

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年2月10日

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  • 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

摘要

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.