Preprint Article Version 1 This version is not peer-reviewed

Whole Mitochondrial DNA Sequencing Using Fecal Samples From Domestic Dogs

Takehito Sugasawa
* ,
Kieu D. M. Nguyen
,
Norihiro Otani
,
Kiyoshi Maehara
,
Fuka Kamiya
,
Atsushi Hirokawa
,
Tohru Takemasa
,
Koichi Watanabe
,
Takeki Nish
,
Ken Sato
,
Suzuka Shimmura
,
Yoichiro Takahashi
,
Yasuharu Kanki
Version 1 : Received: 23 August 2024 / Approved: 26 August 2024 / Online: 27 August 2024 (11:00:32 CEST)

How to cite: Sugasawa, T.; Nguyen, K. D. M.; Otani, N.; Maehara, K.; Kamiya, F.; Hirokawa, A.; Takemasa, T.; Watanabe, K.; Nish, T.; Sato, K.; Shimmura, S.; Takahashi, Y.; Kanki, Y. Whole Mitochondrial DNA Sequencing Using Fecal Samples From Domestic Dogs. Preprints 2024, 2024081864. https://doi.org/10.20944/preprints202408.1864.v1 Sugasawa, T.; Nguyen, K. D. M.; Otani, N.; Maehara, K.; Kamiya, F.; Hirokawa, A.; Takemasa, T.; Watanabe, K.; Nish, T.; Sato, K.; Shimmura, S.; Takahashi, Y.; Kanki, Y. Whole Mitochondrial DNA Sequencing Using Fecal Samples From Domestic Dogs. Preprints 2024, 2024081864. https://doi.org/10.20944/preprints202408.1864.v1

Abstract

The medical care and welfare of domestic dogs are now respected worldwide at the same level as that of humans. We previously established a test method to determine whole mitochondrial DNA (mtDNA) using oral mucosal DNA that may be useful for the medical care and welfare of dogs. However, the sample types tested in dogs are not limited to those obtained from the oral mucosa. Therefore, in the present study, we attempted to establish a test method to determine whole mtDNA sequences using feces, which represents the least invasive specimen. Two Japanese domestic dogs were used in the present study. DNA was extracted from approximately 100 mg of fresh feces from each dog, and PCRs were performed using four primer-pairs that can amplify whole mtDNA. Following PCR, amplicons were pooled to create a DNA library using an experimental robot with an original program. Data were then acquired via NGS and data analysis was performed. The results showed that the whole mtDNA sequence of the two dogs was determined with high accuracy. Our results suggest that feces can be adapted for mitochondrial disease and individual identification testing and could serve as a useful testing method for the future medical care and welfare of domestic dogs.

Keywords

domestic dogs; whole mitochondrial DNA; feces; fecal samples; next-generation sequencer

Subject

Biology and Life Sciences, Life Sciences

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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