Ultralow input genome sequencing library preparation from a single tardigrade specimen

Yuki Yoshida, Sayuri Konno, Ryousuke Nishino, Yumi Murai, Masaru Tomita, Kazuharu Arakawa

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Tardigrades are microscopic animals that enter an ametabolic state called anhydrobiosis when facing desiccation and can return to their original state when water is supplied. The genomic sequencing of microscopic animals such as tardigrades risks bacterial contamination that sometimes leads to erroneous interpretations, for example, regarding the extent of horizontal gene transfer in these animals. Here, we provide an ultralow input method to sequence the genome of the tardigrade, Hypsibius dujardini, from a single specimen. By employing rigorous washing and contaminant exclusion along with an efficient extraction of the 50 ~ 200 pg genomic DNA from a single individual, we constructed a library sequenced with a DNA sequencing instrument. These libraries were highly reproducible and unbiased, and an informatics analysis of the sequenced reads with other H. dujardini genomes showed a minimal amount of contamination. This method can be applied to unculturable tardigrades that could not be sequenced using previous methods.

Original languageEnglish
Article numbere57615
JournalJournal of Visualized Experiments
Volume2018
Issue number137
DOIs
Publication statusPublished - 2018 Jul 15

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Keywords

  • Contamination-free
  • Freeze-thaw homogenization
  • Genetics
  • Genome sequencing
  • Hypsibius dujardini
  • Issue 137
  • Tardigrade
  • Ultralow input

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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