The Method of Eliminating the Wolbachia Endosymbiont Genomes from Insect Samples Prior to a Long-Read Sequencing

Keizo Takasuka, Kazuharu Arakawa

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

When extracting DNA of invertebrates for long-read sequencing, not only enough quantity and size of the DNA but, depending on the species, elimination of contamination of endosymbiotic Wolbachia genome also has to be achieved. These requirements become troublesome, especially in small-sized species with a limited number of individuals available for the experiment. In this chapter, using tiny parasitoid wasps (Reclinervellus nielseni) parasitizing spiders as hosts, we developed a method of eliminating the Wolbachia genomes by means of an antibiotic administration to adult wasps via honey solution. Twenty days of rifampicin treatment since their emergence from cocoons resulted in a significant decrease in the Wolbachia genomes while keeping good DNA conditions for nanopore sequencing. An adequate quantity of DNA was then gained by pooling several individuals. The method could be applied to other insects or invertebrates that can be maintained by laboratory feeding with liquid food.

Original languageEnglish
Pages (from-to)101-112
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Volume2632
DOIs
Publication statusPublished - 2023

Keywords

  • Antibiotics
  • Contamination
  • DNA extraction
  • Endocellular bacterial symbiont
  • Genome
  • Nanopore sequence
  • Rifampicin
  • qPCR

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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