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1

Molecular identification of hairtail species (Pisces: Trichiuridae) based on PCR-RFLP analysis of the mitochondrial 16S rRNA gene

100%
Chakraborty A., Aranishi F., Iwatsuki Y.
Journal of Applied Genetics
|
2005
|
vol. 46
|
issue 4
381-385
EN
A rapid PCR-RFLP analysis was designed to identify 3 closely related species of hairtails: Trichiurus lepturus, T. japonicus, and Trichiurus sp. 2, basing on partial sequence data (600 bp) of the mitochondrial DNA encoding the 16S ribosomal RNA (16S rRNA) gene. Restriction digestion analysis of the unpurified PCR products of these 3 species, using EcoRI and VspI endonucleases, generated reproducible species-specific restriction patterns showing 2 fragments (250 bp and 350 bp) for T. lepturus in EcoRI digestion and 2 fragments (196 bp and 404 bp) for T. japonicus in VspI digestion, whereas no cleavage was observed for Trichiurus sp. 2 in both EcoRI and VspI digestions. The PCR-RFLP technique developed in this study proved to be a rapid, reliable and simple method that enables easy and accurate identification of these 3 closely related species of the genus Trichiurus.
2

Use of cytochrome b polymorphism for species identification of biological material derived from cattle, sheep, goats, roe deer and red deer

100%
Natonek-Wisniewska M., Slota E., Kalisz B.
Folia Biologica
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2010
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vol. 58
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issue 1-2
47-50
EN
The objective of this study was species identification of the following biological trace material: skin, blood stains, meat samples and jawbone with a tooth, which were the subject of expert opinion ordered by a court. The expert appraisement was conducted by an analysis of a cytochrome b fragment. The choice of mtDNA fragment for analysis was based on its conservation in mammals which enabled several farm and wild species to be identified with one pair of primers. The PCR product was differentiated by Tsp509I and AluI enzymes. Due to problems with amplification of roe deer DNA, primers specific to this species only, flanking a cytochrome b fragment (Y14951.1), were designed. On the basis of this analysis, it was concluded that the skin sample was derived from a goat, dried blood from a roe deer, the jawbone fromcattle, and twomeat samples froma roe deer and red deer. od from a roe deer, the jawbone fromcattle, and twomeat samples froma roe deer and red deer. This method allowed rapid and efficient identification of several species of mammals using diverse biological material.
3

Identification of gadoid species (Pisces, Gadidae) by PCR-RFLP analysis

100%
Aranishi F., Okimoto T., Izumi S.
Journal of Applied Genetics
|
2005
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vol. 46
|
issue 1
69-73
EN
A rapid PCR-RFLP analysis was optimized to identify the presence of 3 closely related gadoid fish species: Alaska pollack Theragra chalcogramma, Pacific cod Gadus macrocephalus and Atlantic cod Gadus morhua in commercial seafood products. Gadoid universal primers were designed for PCR amplification of a 558-bp fragment encoding the mitochondrial cytochrome b gene. Without purification of the PCR products, double digestion with Eco32I and Eco105I restriction enzymes generated reproducible species-specific restriction patterns visualizing 3 fragments (106 bp, 161 bp and 291 bp) in Alaska pollack and 2 fragments (106 bp and 452 bp) in Pacific cod, whereas no cleavage was observed in Atlantic cod. This PCR-RFLP analysis is simple, rapid and reliable, and therefore can be routinely applied to discover fraudulent substitution among 3 economically important gadoid species in commercial seafood products.
4

Restriction fragment length polymorphism of mitochondrial DNA and phylogenetic relationships among five species of Indian freshwater turtles

75%
Rohilla M. S., Tiwari P. K.
Journal of Applied Genetics
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2008
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vol. 49
|
issue 2
167-182
EN
DNA-based identification of species for phylogenetic analysis as well as forensic identification is widely being carried out with the help of polymerase chain reaction (PCR). In this study, a successful effort has been made to identify 5 species of Indian freshwater turtles, including 3 hard-shell turtles (Geoemydidae), i.e. Kachuga dhongoka, K. kachuga and Geoclemys hamiltoni, and 2 species of soft-shell turtles (Trionychidae), i.e. Aspideretes gangeticus and Lissemys punctata punctata, by using a well-optimized PCR-RFLP method. The analysis of nucleotide sequence variations in the PCR-amplified mitochondrial cyt-b genes (encoding cytochrome b) from the 5 species revealed its usefulness in the taxonomic differentiation of these species. On the basis of cyt-b sequence data and the PCR-RFLP pattern, a phylogeny was developed to resolve the genetic relationships between these species, living in the same habitat type. In comparison, the PCR-RFLP of mitochondrial 16S rDNA genes appeared less decisive in analysing phylogenetic relationships or even in species differentiation. Further, the molecular method (PCR-RFLP) developed here is simple, rapid, reliable and reproducible; hence it can be routinely applied for species identification, essential for conservation and management of endangered chelonian species.
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