Ultralow input genome sequencing library preparation from a single tardigrade specimen

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

研究成果: Article査読

2 被引用数 (Scopus)

抄録

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.

本文言語English
論文番号e57615
ジャーナルJournal of Visualized Experiments
2018
137
DOI
出版ステータスPublished - 2018 7 15

ASJC Scopus subject areas

  • 神経科学(全般)
  • 化学工学(全般)
  • 生化学、遺伝学、分子生物学(全般)
  • 免疫学および微生物学(全般)

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