Pretreatment method for DNA barcoding to analyze gut contents of rotifers

Hye Ji Oh, Paul Henning Krogh, Hyun Gi Jeong, Gea Jae Joo, Ihn Sil Kwak, Sun Jin Hwang, Jeong Soo Gim, Kwang Hyeon Chang, Hyunbin Jo

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

We designed an experiment to analyze the gut content of Rotifera based on DNA barcoding and tested it on Asplanchna sp. in order to ensure that the DNA extracted from the rotifer species is from the food sources within the gut. We selected ethanol fixation (60%) to minimize the inflow effects of treated chemicals, and commercial bleach (the final concentration of 2.5%, for 210 s) to eliminate the extracellular DNA without damage to the lorica. Rotifers have different lorica structures and thicknesses. Therefore, we chose a pretreatment method based on Asplanchna sp., which is known to have weak durability. When we used the determined method on a reservoir water sample, we confirmed that the DNA fragments of Chlorophyceae, Diatomea, Cyanobacteria, and Ciliophora were removed. Given this result, Diatomea and cyanobacteria, detected from Asplanchna, can be considered as gut contents. However, bacteria were not removed by bleach, thus there was still insufficient information. Since the results of applying commercial bleach to rotifer species confirmed that pretreatment worked effectively for some species of rotifers food sources, in further studies, it is believed to be applicable to the gut contents analysis of more diverse rotifers species and better DNA analysis techniques by supplementing more rigorous limitations.

Original languageEnglish
Article number1064
JournalApplied Sciences (Switzerland)
Volume10
Issue number3
DOIs
Publication statusPublished - 1 Feb 2020

Bibliographical note

Publisher Copyright:
© 2020 by the author.

Keywords

  • Commercial bleach
  • Eliminate the extracellular DNA
  • Gut content of Rotifera
  • Pretreatment

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