Amplificação isotérmica mediada por loop para identificação de espécies puras e híbridos de peixe da família Serrasalmidae utilizados na piscicultura brasileira

• Autor(a) principal: Santos, Natalia dos
• Data de Publicação: 2022
• Tipo de documento: Trabalho de conclusão de curso
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/217745
• topic: LAMP; Fishes; Genetic analysis; Conservation; Análise genética; Peixes

Loop-mediated isothermal amplification (LAMP) is a molecular technique that has been widely used in several biological fields, due to the ease of application in species identification. The design of core and loop primers for using the amplification technique is more complex than PCR primers, as there are four core primers and two loop specific that recognize six distinct sequences of the target DNA for amplification, in contrast with only two of the normal PCR. However, this technique makes it possible to identify species quickly, accurately and at lower cost when compared to PCR. In aquaculture, the identification of parental species and their respective hybrids through LAMP allows the diagnosis and objective analysis that can be used to adequately manage fish farms, in the identification of species at the beginning of the cultivation, and allows increased knowledge regarding impacts environment impacts caused by cultivated species, affecting wild populations. The objective of this work was to apply and standardize the LAMP technique for the identification of pure species of Colossoma macropomum, Piaractus mesopotamicus and its hybrid (“tambacu”), thus enabling faster, more accurate, efficient and low-cost analysis, with quick visualization, in a simple and objective manner. It was possible to develop for the first time a LAMP protocol for the diagnosis of P. mesopotamicus and C. macropomum species, as well as for tambacu, using the RAG2 gene. It was also possible to validate the LAMP technique to identify the species of the fish in question, diagnosing and differentiating pure and hybrid individuals.