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A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS

BACKGROUND: Concern has been raised about DNA extraction from Mycobacterium tuberculosis due to its complex procedure. This study demonstrates a simple and fast DNA extraction method of mycobacterial genome to subsequent molecular investigation, such as Polymerization Chain Reaction (PCR) amplificat...

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Autores principales: Koentjoro, Maharani Pertiwi, Donastin, Adyan, Prasetyo, Endry Nugroho
Formato: Online Artículo Texto
Lenguaje:English
Publicado: African Traditional Herbal Medicine Supporters Initiative (ATHMSI) 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8457349/
https://www.ncbi.nlm.nih.gov/pubmed/34595382
http://dx.doi.org/10.21010/ajidv15i2S:2
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author Koentjoro, Maharani Pertiwi
Donastin, Adyan
Prasetyo, Endry Nugroho
author_facet Koentjoro, Maharani Pertiwi
Donastin, Adyan
Prasetyo, Endry Nugroho
author_sort Koentjoro, Maharani Pertiwi
collection PubMed
description BACKGROUND: Concern has been raised about DNA extraction from Mycobacterium tuberculosis due to its complex procedure. This study demonstrates a simple and fast DNA extraction method of mycobacterial genome to subsequent molecular investigation, such as Polymerization Chain Reaction (PCR) amplification, with species-specific primers and sequencing. MATERIALS AND METHODS: Total DNA was isolated from M. tuberculosis cultured by using boil method. DNA was evaluated via measures of DNA quantity and quality (absorbance at 230, 260 and 280 nm), DNA integrity (electrophoresis). Molecular tests were tested namely PCR and sequencing. CONCLUSIONS: The quality of DNA obtained is acceptable for PCR and sequencing analysis. These findings demonstrate that the method used is inexpensive and suitable for minimum infrastructure facilities.
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spelling pubmed-84573492021-09-29 A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS Koentjoro, Maharani Pertiwi Donastin, Adyan Prasetyo, Endry Nugroho Afr J Infect Dis Article BACKGROUND: Concern has been raised about DNA extraction from Mycobacterium tuberculosis due to its complex procedure. This study demonstrates a simple and fast DNA extraction method of mycobacterial genome to subsequent molecular investigation, such as Polymerization Chain Reaction (PCR) amplification, with species-specific primers and sequencing. MATERIALS AND METHODS: Total DNA was isolated from M. tuberculosis cultured by using boil method. DNA was evaluated via measures of DNA quantity and quality (absorbance at 230, 260 and 280 nm), DNA integrity (electrophoresis). Molecular tests were tested namely PCR and sequencing. CONCLUSIONS: The quality of DNA obtained is acceptable for PCR and sequencing analysis. These findings demonstrate that the method used is inexpensive and suitable for minimum infrastructure facilities. African Traditional Herbal Medicine Supporters Initiative (ATHMSI) 2021-09-01 /pmc/articles/PMC8457349/ /pubmed/34595382 http://dx.doi.org/10.21010/ajidv15i2S:2 Text en Copyright: © 2021 Afr. J. Infect. Diseases https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License
spellingShingle Article
Koentjoro, Maharani Pertiwi
Donastin, Adyan
Prasetyo, Endry Nugroho
A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS
title A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS
title_full A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS
title_fullStr A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS
title_full_unstemmed A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS
title_short A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS
title_sort simple method of dna extraction of mycobacterium tuberculosis from sputum cultures for sequencing analysis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8457349/
https://www.ncbi.nlm.nih.gov/pubmed/34595382
http://dx.doi.org/10.21010/ajidv15i2S:2
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