DNA Extraction of Cynopterus brachyotis of Labuhan Ratu VII Based on Column Method in Supporting its Molecular Species Confirmation
Article Sidebar
Main Article Content
Abstract
Background. Bats (Chiroptera) are true flying mammals and nocturnal. Ecologically, bats play important roles as pollinators, seed dispersers, guano producers, biocontrol, and hosts a range of viruses. Cynopterus brachyotis belongs to fruit-eating bats and is considered morphologically cryptic with C. sphinx.
Aims. To support morphological identification, molecular species confirmation needed to be conducted. DNA extraction, as an initial step, plays a crucial role in molecular analysis.
Methods. The quality and quantity of extracted DNA determine the success rate of sequencing for species confirmation in bats. In this study, oropharyngeal samples were used.
Result. The research procedures included bat capture and DNA oropharyngeal swab sample collection, sample preparation, DNA extraction using a silica column-based commercial kit, DNA concentration measurement using a Qubit assay, and electrophoresis.
Conclusion. The Qubit assay showed DNA concentrations of >1 ng/µL, while electrophoresis did not reveal bright DNA bands.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Al-Shuhaib, M. B., and Hashim, H. O. (2023). Mastering DNA chromatogram analysis In Sanger sequencing for reliable clinical Analysis. Journal of Genetic Engineering and Biotechnology, 21(115): 1-12.
Arnaout, Y., Djelouadji, Z., Robardet, E., Cappelle, J., Cliquet, F., Touzalin, F., Jimenez, G., Hurstel, S., Borel, C., & Picard-Meyer, E. (2022). Genetic identification of bat species for pathogen surveillance across France. PLoS ONE, 17(1), e0261344.
Bruijns, B., Hoekema, T., Oomens, L., Tiggelaar, R., and Gardeniers, H. (2022). Performance of Spectrophotometric and Fluorometric DNA Quantification Methods. Analytica, 3(3), 371-384.
Guan, X., Britzke, E. R., Piaggio, A. J., Bergman, D. L., Van Pelt, L., & Lance, R. F. (2020). Genetic assays for guano-based identification of species and sex in bats of the United States and Canada. Journal of Mammalogy, 101(4), 970–978.
Hughes, M., Lennon, R., & Fenton, M. B. (2024). Successful DNA amplification from non-destructive buccal swabbing in Vespertilionid and Rhinolophid bats. Conservation Genetics Resources. 16, 1–7.
Lam, T. T. H., Dinh, Q. M., Nguyen, T. H. D., Phan, G. H. 2024. Genetic Diversity ad Phylogenetic Tree of GrainGenus in Viatnamese Mekong Delta Based on Mitochondrial Cytochrome B Gene Sequence. Egyptian Journal of Aquatic Research, 50: 498-505.
Manek, Y., Elu, A., Hendrik, A. C., Blegur, W. A., and Bullu, N. I. (2020). Identification of Types and Morphometric Characteristics of Bats in Fatubaun Cave, Manufui Village, Santian District, South Central Timor Regency. Journal of Dry Land Science, 3(2): 42-46.
Marlinda, D., Mailani, C., and Kamal, S 2019. Identification of Bat Species (Chiroptera) in the Campus Area of UIN Ar-Raniry Banda Aceh. Proceedings of the National Seminar on Biotics, 79-82.
Morcatty, T. Q., Pereyra, P. E., Ardiansyah, A., Imron, M. A., Hedger, K., Campera, M., and Nijiman, V. 2022. Risk of Viral Infectious Disease from Live Bats, Primates, Rodents and Carnivores for Sale in Indonesian Wildlife Markets. Viruses, 14(12): 2756-2758.
Mubarok, H., Handayani, N. S. N., Maryanto, I., and Arisuryanti, A. (2023). Phylogenetic and Genetic Variation Analysis of Lesser Short-Nosed Fruit Bat Cynopterus Brachyotis (Müller 1838) on Java Island, Indonesia, Inferred from Mitochondrial D Loop. Journal of Genetic Engineering and Biotechnology, 21(1): 1-10.
Pearce, D. L., Edson, J. E., Jenelle, C. S., and Walter, W. D. 2024. Evaluation of DNA yield from various tissue and sampling sources for use in single nucleotide polymorphism panels. Scientific Reports, 14(11340): 1-8.
Ramona, F. (2019). Diversity and Potential of Megachiroptera Bats as Dispersers and Pollinators in Harapan Forest, Jambi. Journal Bio-Site, 4(1): 1-11.
Ransaleleh, T. A., Kawatu, M., Wahyuni, I., Onibala, J., and Kambey, P. (2024). Morphology of bats (Chiroptera: Megachiroptera) around the plantation of Sawangan Village, North Minahasa. Zootec Journal, 44(1): 95-106.
Safitri, Z., Prayogo, H., and Erianto. (2020). Diversity of Bat Species (Chiroptera) in the Tanjungpura University Area, Pontianak City. Journal of Sustainable Forests, 8(2): 429-440.
Socea, J. N., Stone, V. N., Qian, X., Gibbs, P. L., and Levinson, K. J. (2023). Implementing laboratory automation for next-generation sequencing: benefits and challenges for library preparation. Front. Public Health 11:1195581.
Tangguda, S., Valentine, R. Y., Hariyadi, D. R., and Sudiarsa, I. N. 2022. Utilization of Bat Droppings as Guano Fertilizer in Bolok Village, West Kupang, East Nusa Tenggara. Journal of Agriculture, 33(3): 289-295.
Triasih, D., Dewi, R. R., Erwanto, Y., and Fitrianto, N. A. 2020. Comparison of Isolation Methods on the Detection of Cowhide, Buffalo, Goat, and Pig Skins as a Raw Material for Skin Patches. Triton Journal, 11(1): 37-44.
Walker FM, Tobin A, Simmons NB, Sobek CJ, Sanchez DE, Chambers CL, et al. (2019) A fecal sequel: Testing the limits of a genetic assay for bat species identification. PLoS sONE, 14(11): e0224969.