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

Authors

  • Maharani Pertiwi Koentjoro 1Medical Laboratory Technology, Faculty of Health, Universitas Nahdlatul Ulama Surabaya, Jl. Jemursari No. 57, Surabaya, Indonesia
  • Adyan Donastin 2Faculty of Medicine, Universitas Nahdlatul Ulama Surabaya, Jl. Jemursari No. 57, Surabaya, Indonesia
  • Endry Nugroho Prasetyo 3Departement of Biology, Faculty of Science and Data Analytics, Institut Teknologi Sepuluh Nopember, Gedung H, Kampus ITS Sukolilo, Surabaya, Indonesia

DOI:

https://doi.org/10.21010/Ajid%20v15i2S.2

Keywords:

DNA extraction, Mycobacterium tuberculosis, sequencing, sputum

Abstract

 

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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Published

2021-09-01

How to Cite

Koentjoro, M. P., Donastin, A., & Prasetyo, E. N. (2021). A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS. African Journal of Infectious Diseases (AJID), 15(2 Suppleme), 19–22. https://doi.org/10.21010/Ajid v15i2S.2