Publicação
Comparative Analysis of DNA Extraction Methods for Individual Varroa destructor
| Resumo: | The ectoparasitic mite Varroa destructor is one of the major honey bee threats and it is associated to population worldwide decline. Genetic analyses using the mtDNA of V. destructor are fundamental for establishing the taxonomy and distribution of the mites. Consequently, low-quality DNA can lead to inaccurate or inconsistent data, making genetic interpretation more challenging. In this study, we compared the concentration and quality of DNA extracted from individual female V. destructor using two different commercial kits, aiming to identify the optimal method for obtaining high-quality DNA. Total DNA was extracted from mites using both an automated and manual extracted method. In addition, manual kit extraction tested three incubation procedure (1h, 5h, and overnight). DNA concentration was quantified using three different instruments: the SpectroStarVR Nano LVis Plate spectrophotometer, the Quantus™ Fluorometer apparatus, and NanoDrop™. The manual extraction DNA concentration did not vary across incubation times and the concentration values varied between 0.240-0.545 ng/μl (Quantus), 0.72-4.49 ng/μl (spectrophotometer), and 0.0-1.47 ng/μl (NanoDrop). While extraction automatic approach yielded higher respectively 0.483-0.631 ng/μl, 20.33-9.0 ng/μl, and 5.3-6.8 ng/μl. In conclusion, the automated kit extraction seems to be the best extraction method since it produced the higher-concentration DNA using only one individual mite. |
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| Autores principais: | Costa, Maíra |
| Outros Autores: | Lopes, Ana; Yadró Garcia, Carlos A.; Vitrio, Nathalia; Gonçalves, Telma; Pinto, M. Alice; Henriques, Dora |
| Assunto: | DNA extraction DNA quantification DNA quality Varroa destructor Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology |
| Ano: | 2024 |
| País: | Portugal |
| Tipo de documento: | documento de conferência |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Instituto Politécnico de Bragança |
| Idioma: | inglês |
| Origem: | Biblioteca Digital do IPB |
| Resumo: | The ectoparasitic mite Varroa destructor is one of the major honey bee threats and it is associated to population worldwide decline. Genetic analyses using the mtDNA of V. destructor are fundamental for establishing the taxonomy and distribution of the mites. Consequently, low-quality DNA can lead to inaccurate or inconsistent data, making genetic interpretation more challenging. In this study, we compared the concentration and quality of DNA extracted from individual female V. destructor using two different commercial kits, aiming to identify the optimal method for obtaining high-quality DNA. Total DNA was extracted from mites using both an automated and manual extracted method. In addition, manual kit extraction tested three incubation procedure (1h, 5h, and overnight). DNA concentration was quantified using three different instruments: the SpectroStarVR Nano LVis Plate spectrophotometer, the Quantus™ Fluorometer apparatus, and NanoDrop™. The manual extraction DNA concentration did not vary across incubation times and the concentration values varied between 0.240-0.545 ng/μl (Quantus), 0.72-4.49 ng/μl (spectrophotometer), and 0.0-1.47 ng/μl (NanoDrop). While extraction automatic approach yielded higher respectively 0.483-0.631 ng/μl, 20.33-9.0 ng/μl, and 5.3-6.8 ng/μl. In conclusion, the automated kit extraction seems to be the best extraction method since it produced the higher-concentration DNA using only one individual mite. |
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