Species Identification of Red Deer (Cervus elaphus), Roe Deer (Capreolus capreolus), and Water Deer (Hydropotes inermis) Using Capillary Electrophoresis-Based Multiplex PCR
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Preparation
2.2. DNA Extraction
2.3. Primer Design
2.4. Single and Multiplex PCR Conditions
2.5. Specificity and Sensitivity of Multiplex PCR
3. Results and Discussion
3.1. The Specificity of Newly Designed Species-Specific Primers
3.2. Specificity and Sensitivity of Capillary Electrophoresis-Based Multiplex PCR
3.3. Application and Validation of Capillary Electrophoresis-Based Multiplex PCR Using Meat DNA Mixtures
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Target Species | Primer Name | Sequences (5′ → 3′) | Target Genes | Amplicon Size (bp) | Accession No. |
---|---|---|---|---|---|
Red deer | Deer 2 F | TGGACAACTAGCATCTGTCT | cyt b | 79 | JF489133.1 |
Deer 2 R | GAGGTTGTTTTCGATTGTGCTGGTG | ||||
Roe deer | Roe deer 3 F | CAGCCTTCCTATTAACCCCT | 12 S rRNA | 126 | KJ681490.1 |
Roe deer 3 R | AGGTGTCATGAGCTACAGGC | ||||
Water deer | Water deer 1 F | CATGATTCAACCCTACAATTC | D-loop | 160 | NC011821 |
Water deer 1 R | GGCGCTTAAATACATACCTTGCT |
The Ratio of Meat DNA Mixtures (%) | |||
---|---|---|---|
Red Deer | Roe Deer | Water Deer | |
Roe deer and water deer in red deer | 80 | 10 | 10 |
90 | 5 | 5 | |
98 | 1 | 1 | |
99 | 0.5 | 0.5 | |
99.8 | 0.1 | 0.1 | |
Red deer and roe deer in water deer | 10 | 10 | 80 |
5 | 5 | 90 | |
1 | 1 | 98 | |
0.5 | 0.5 | 99 | |
0.1 | 0.1 | 99.8 |
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Kim, M.-J.; Lee, Y.-M.; Suh, S.-M.; Kim, H.-Y. Species Identification of Red Deer (Cervus elaphus), Roe Deer (Capreolus capreolus), and Water Deer (Hydropotes inermis) Using Capillary Electrophoresis-Based Multiplex PCR. Foods 2020, 9, 982. https://doi.org/10.3390/foods9080982
Kim M-J, Lee Y-M, Suh S-M, Kim H-Y. Species Identification of Red Deer (Cervus elaphus), Roe Deer (Capreolus capreolus), and Water Deer (Hydropotes inermis) Using Capillary Electrophoresis-Based Multiplex PCR. Foods. 2020; 9(8):982. https://doi.org/10.3390/foods9080982
Chicago/Turabian StyleKim, Mi-Ju, Yu-Min Lee, Seung-Man Suh, and Hae-Yeong Kim. 2020. "Species Identification of Red Deer (Cervus elaphus), Roe Deer (Capreolus capreolus), and Water Deer (Hydropotes inermis) Using Capillary Electrophoresis-Based Multiplex PCR" Foods 9, no. 8: 982. https://doi.org/10.3390/foods9080982