Genetic divergence among three morphs of Acentrogobius pflaumii (Gobiidae) around Japan and their identification using multiplex haplotype-specific PCR of mitochondrial DNA

Melt-curve-multiplex-haplotype-specific-PCR, a valuable tool for biological studies: Application in congeneric species discrimination assay

“…during digestion in stomach contents or food processing) and samples of early life stages of fish species (Gil, 2007;Hyde et al, 2005;Teletchea, 2009). To overcome these difficulties, many DNA-based discrimination assays have been developed for commercial fish-product controls (Michelini et al, 2007;Trotta et al, 2005), for the identification of "cryptic species" or congeneric juveniles (Apostolidis et al, 2008;Matsui et al, 2012) and also for studies of fish eggs and larvae (Hyde et al, 2005;Rocha-Olivares, 1998). …”

“…Particularly the use of species-specific primers is considered an advantageous method for the routine analysis of numerous samples, especially in discrimination studies that include small number of species (Hyde et al, 2005;Michelini et al, 2007). Therefore, several assays with species-specific primers have been applied in biological and ecological studies (Ma et al, 2012;Matsui et al, 2012;Piergiorge et al, 2014).…”

Distribution and habitat use of three Acentrogobius (Perciformes: Gobiidae) species in the coastal waters of Japan

“…C. These three species were characterized, respectively, as follows (Suzuki et al 2004): no predorsal scales and no dark spot on the first dorsal fin; dense predorsal scales and a dark spot on the first dorsal fin; and predorsal scales clearly confirmed (judging from their photo) and a large brownish oval spot on the caudalfin base. Additionally, Matsui et al (2012) showed that these morphs are genetically divergent, which supports the validity of their classification as distinct species.…”

“…B, and Acentrogobius sp. C. sensu Suzuki et al (2004) and Matsui et al (2012)] and to clarify their habitat use across the Japanese archipelago.…”

Mitochondrial DNA sequence divergence between “Kunimasu” Oncorhynchus kawamurae and “Himemasu” O. nerka in Lake Saiko, Yamanashi Prefecture, Japan, and their identification using multiplex haplotype-specific PCR

“…1040 bp in Himemasu were predicted to be amplified. For internal positive control of the PCR conditions, a pair of primers expected to commonly anneal to either of the two species, L-M-12S; 5'-ACA AAC TGG GAT TAG ATA CCC CAC TAT GCC-3' and H-A-16S; 5'-GTT TTT GGT AAA CAC GCG AGG CTT ATG TTT GCC GAG-3' (Matsui et al 2012), which are complements of the primers of Kawaguchi et al (2001), producing a ca. 1,500 bp amplicon, were also included in MHS-PCR.…”