Bacterial genomic dna extraction: a comparison of commercial and in house methods

• Autor(a) principal: Bierhals, Nayanna Dias
• Data de Publicação: 2020
• Outros Autores: Brixner, Betina, Silva, Karoline Schroder da, Oliveira, Caio Fernando de, Renner, Jane Dagmar Pollo
• Tipo de documento: Artigo
• Idioma: por
• Título da fonte: Saúde (Santa Maria)
• Texto Completo: https://periodicos.ufsm.br/revistasaude/article/view/41122
• topic: Bacterial DNA; Real time PCR; Gram-positive bacteria; Gram-negative bacteria.; DNA bacteriano; PCR em tempo real; Bactérias Gram-negativas; Bactérias Gram-positivas.

Objective: To compare and evaluate four different methods of bacterial DNA extraction. Methods: We use control strains of Gram-negative and Gram-positive bacteria for DNA extraction by the following methods: boiling, alkaline lysis, in house kit and commercial kit. We compare the results of quantitative and qualitative analyses, real time PCR efficiency, time, reproducibility and cost. On quantitative and qualitative analyses we observe some variations in results obtained for each strain and method, however, boiling, alkaline lysis and in house kit showed similar purity. Real time PCR results confirmed the efficiency of all methods tested. Results: Regarding time, reproducibility and cost the boiling method stood out with good results. Conclusion: The alkaline lysis method achieved better performance presenting excellent PCR amplifications, good DNA amounts, satisfactory purity, ideal time and cost and reproducibility acceptable for both Gram-positive and Gram-negative bacteria, being a very interesting option for application on laboratory routine.