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1

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

100%
Aranishi F., Okimoto T., Izumi S.
Journal of Applied Genetics
|
2005
|
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.
2

Paramecium sexaurelia-intra-specific polymorphism and relationships with other P. aurelia spp, revealed by cytochrome b sequence data

100%
Przybos E., Barth D., Berendonk T. U.
Folia Biologica
|
2010
|
vol. 58
|
issue 1-2
55-60
EN
Genetic distances among strains of P. sexaurelia were compared by sequencing the mitochondrial cytochrome b gene. The relationships of P. sexaurelia with representative strains of other species of the P. aurelia complex and related species, i.e. P. schewiakoffi, P. jenningsi and P. multimicronucleatum were evaluated. All investigated P. sexaurelia and P. dodecaurelia strains grouped together with high support. This P. sexaurelia/P. dodecaurelia cluster was furthermore composed of three distinct, strongly supported subgroups. Two of these groups contained both P. sexaurelia and P. dodecaurelia strains, suggesting that these species are not monophyletic. The third branch was composed of strains from Sevilla, Spain and was deeply separated (12-14 % p-distance) from the other P. sexaurelia/P. dodecaurelia clades. This illustrates the urgent need for further work and more intense sampling of these .rare. species, in order to understand the status and the relationships of P. sexaurelia and P. dodecaurelia within the P. aurelia species complex. We recommend that general investigations on the speciation process be conducted within and between species of the P. aurelia complex due to the high genetic variation combined with observations that for some of the species possible cryptic speciation may have occurred. This is emphasized by data showing that for some species within this complex, species status may be less static and more dynamic than originally thought.
3

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
|
2010
|
vol. 58
|
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.
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
|
2008
|
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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