RT info:eu-repo/semantics/masterThesis
T1 Development of a hands-on genomics educational tool using mitochondrial DNA PCR-free enrichment and MinION-based species apportionment
A1 Gonzalez Navasa, Carolina
A2 Máster Universitario en Biomedicina Por la Universidad de la Laguna
K1 biomedicina
AB Identification of different species can be performed attending to the variation in theDNA sequences. Mitochondrial DNA (mtDNA) constitutes the ideal model as itprovides an opportunity for eukaryotic classification as it is available from a largenumber of metazoan species. We aimed to devise a protocol to allow the sequencing ofthe mtDNA without using PCR with cutting-edge third-generation next generationsequencing (NGS) technology to identify eukaryotic organisms. Two procedures wereassessed for mtDNA enrichment from distinct food products: the commercial solutionQIAprep Spin Miniprep Kit (QIAGEN), and the home-made protocol of classic alkalinelysis for plasmid isolation. The enrichment of mtDNA versus nuclear DNA wasevaluated by quantitative real-time PCR and short-read sequencing. As a proof ofconcept, a mock sample with the DNAs from the best performing enrichment protocolwas finally processed with MinION (Oxford Nanopore Technologies) and NanoDJ toallow a qualitative identification of the food components. While the protocols still needimprovements to facilitate sample processing, we envisage this hands-on experience asa learning-by-doing instrument for transdisciplinary education adapted to basiclaboratory resource settings.Keywords: Nanopore sequencing; educational tool; genomics; species classification
YR 2018
FD 2018
LK http://riull.ull.es/xmlui/handle/915/11538
UL http://riull.ull.es/xmlui/handle/915/11538
LA es
DS Repositorio institucional de la Universidad de La Laguna
RD 27-dic-2024