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| Buddhakosai, W.; Sukmak, M.; Klinsawat, W.; Duangchantrasiri, S. | |
| Discrimination of tiger using a novel high resolution melting (HRM) and multiplex SNP-specific HRM (MSS-HRM) technique | |
| 2014 Forensic Science International: Genetics (13): 30-33 | |
The discrimination of tiger subspecies or populations can facilitate their conservation by tracking this illegal trade. One recent molecular method has been developed for discriminating tiger subspecies using mitochondrial SNP data [1]. The method requires capillary electrophoresis, which may not be readily available in some laboratories. We therefore have designed and validated an alternative method based on high resolution melting (HRM) analysis; single HRM and novel multiplex SNP-specific HRM (MSS-HRM), for discrimination of tiger DNA, specifically the Bengal, Amur, Indochinese, Malayan tiger and new interesting haplotype in the major site of Thailand (WEFCOM: Western Forest Complex) [2]. The HRM technique is a real-time PCR based methods and relies on the different melting temperature of small amplicons [3]. Differences in nucleotide sequence, amplicon length and GC content all produce different melting temperatures [3-6]. This technique has been used extensively in several studies, including species identification [7,8] and genetic diversity [9,10]. We used 16 sequenced tiger DNA templates: 5 Indochinese, 3 Bengal, 4 Amur, 2 Malayan (haplotype COR4 and COR7), and 2 WEFCOM, as controls (GenBank: KC879287-KC879297, JX477683, KJ508412-KJ508413). |
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