Establishing a Sequencing Method for the Whole Mitochondrial DNA of Domestic Dogs
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Research Ethics
2.2. Dogs and Collected Specimens
2.3. Extraction of Oral Mucosa DNA
2.4. Design of Primer Pairs for mtDNA
2.5. Conditions of the LR-PCR
2.6. Library Preparation of NGS
2.7. NGS Rum
2.8. Bioinformatics Analysis
3. Results
3.1. Oral Mucosal DNA Was Successfully Extracted from and Purified in All Samples
3.2. Optimal LR-PCR Condition Was Determined, and the High Specificity of Each Primer Pair Was Confirmed
3.3. Whole Mitochondrial DNA Was Accurately Resequenced in All Dogs
3.4. Phylogenetic Tree Analysis Confirmed the Similarity among Dogs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Dog No. | Dog Breed/Gender | Total Length (bp) of mtDNA | Number of Variants | Coverage of Variants | Number of Non-Synonymous Amino Acids | Pathogenic Variant | ||
---|---|---|---|---|---|---|---|---|
1 | Kooikerhondje/female | 16,730 | SNV | 101 | Total: 104 | 6135 to 51,966 (Average: 20,178) | 17 | N/A |
MNV | 1 | |||||||
Insertion | 2 | |||||||
Deletion | 0 | |||||||
2 | Keeshond/ female | 16,732 | SNV | 33 | Total: 36 | 6525 to 24,492 (Average: 13,442) | 4 | N/A |
MNV | 0 | |||||||
Insertion | 3 | |||||||
Deletion | 0 | |||||||
3 | Keeshond/ female | 16,732 | SNV | 33 | Total: 36 | 7978 to 33,320 (Average: 17,579) | 4 | N/A |
MNV | 0 | |||||||
Insertion | 3 | |||||||
Deletion | 0 | |||||||
4 | Shiba Inu/ female | 16,730 | SNV | 105 | Total: 109 | 6300 to 45,580 (Average: 17,999) | 20 | N/A |
MNV | 0 | |||||||
Insertion | 3 | |||||||
Deletion | 1 | |||||||
5 | Shiba Inu/ male | 16,730 | SNV | 23 | Total: 25 | 12,637 to 36,862 (Average: 21,158) | 3 | N/A |
MNV | 0 | |||||||
Insertion | 2 | |||||||
Deletion | 0 | |||||||
6 | Beagle/ male | 16,730 | SNV | 21 | Total: 23 | 9690 to 44,299 (Average: 22,209) | 3 | N/A |
MNV | 0 | |||||||
Insertion | 2 | |||||||
Deletion | 0 | |||||||
P.C. | Toy Poodle/ male | 16,730 | SNV | 27 | Total: 29 | 9122 to 28,966 (Average: 17,035) | 3 | N/A |
MNV | 0 | |||||||
Insertion | 2 | |||||||
Deletion | 0 |
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Sugasawa, T.; Matsumoto, Y.; Fang, H.; Takemasa, T.; Komine, R.; Tamai, S.; Gu, W.; Tanaka, K.; Kanki, Y.; Takahashi, Y. Establishing a Sequencing Method for the Whole Mitochondrial DNA of Domestic Dogs. Animals 2023, 13, 2332. https://doi.org/10.3390/ani13142332
Sugasawa T, Matsumoto Y, Fang H, Takemasa T, Komine R, Tamai S, Gu W, Tanaka K, Kanki Y, Takahashi Y. Establishing a Sequencing Method for the Whole Mitochondrial DNA of Domestic Dogs. Animals. 2023; 13(14):2332. https://doi.org/10.3390/ani13142332
Chicago/Turabian StyleSugasawa, Takehito, Yuki Matsumoto, Hui Fang, Tohru Takemasa, Ritsuko Komine, Shinsuke Tamai, Wenchao Gu, Kei Tanaka, Yasuharu Kanki, and Yoichiro Takahashi. 2023. "Establishing a Sequencing Method for the Whole Mitochondrial DNA of Domestic Dogs" Animals 13, no. 14: 2332. https://doi.org/10.3390/ani13142332
APA StyleSugasawa, T., Matsumoto, Y., Fang, H., Takemasa, T., Komine, R., Tamai, S., Gu, W., Tanaka, K., Kanki, Y., & Takahashi, Y. (2023). Establishing a Sequencing Method for the Whole Mitochondrial DNA of Domestic Dogs. Animals, 13(14), 2332. https://doi.org/10.3390/ani13142332