Abstract
Here we present a multiplex PCR assay to distinguish three widespread brown frog species, Rana temporaria, R. arvalis and R. amurensis, based on amplified fragment length. The system consists of four primers to the mitochondrial cytochrome c oxidase subunit I gene, with one universal reverse primer and three species-specific forward primers. Therefore, the PCR gives a fragment of a known specific length, which can be identified by electrophoresis in standard agarose gel. The efficiency of the developed assay was confirmed using both pre-identified and unidentified samples.
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References
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Funding
The study was supported by the Russian Science Foundation (grant no. 21-74-10107).
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ES conceived and designed the study. OE designed the assay and primers. AS, AL and ES conducted the fieldwork. PC and AL conducted the experiments. PC and ES analyzed the data. PC wrote the initial draft of the manuscript. All authors discussed the results, contributed to the draft, read the final manuscript, and gave approval for publication.
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The authors declare no competing interests.
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Ethical approval from Biomedical Ethic Committee of Tyumen State University (record no. 4\15.12.2021).
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NA (no human participants in the study).
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Chernigova, P., Ermakov, O., Lisachev, A. et al. Multiplex PCR assay to distinguish among three widespread brown frog species. Conservation Genet Resour 16, 7–9 (2024). https://doi.org/10.1007/s12686-023-01338-y
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DOI: https://doi.org/10.1007/s12686-023-01338-y