Whole-Genome Deoxyribonucleic Acid Extraction from Mycobacterium Species via the Cetyltrimethylammonium Bromide...

Efficient and reliable DNA extraction is a fundamental step in molecular biology, providing high-quality genetic material for downstream applications such as PCR, sequencing, and genomic analyses. However, the unique cell wall composition of mycobacteria, rich in lipids and mycolic acids, poses significant challenges for efficient DNA isolation. The cetyltrimethylammonium bromide (CTAB) method is widely employed due to its efficiency in extracting high-purity DNA from mycobacterial species, including both fast- and slow-growing strains. The technique involves enzymatic disruption of bacterial cells using lysozyme and proteinase K, followed by the lysis of cell components using CTAB, which selectively binds and forms a complex with polysaccharides and proteins. DNA purification and concentration are achieved by organic solvent extraction and ethanol precipitation. According to our data, the CTAB protocol yields high quantities of DNA (200 - 1000 ng/µL) with high quality and minimal contamination as proven by gel electrophoresis and NanoDrop analysis. The DNA extracted from different mycobacterial species is of high purity and is suitable for various molecular studies. This study demonstrates the CTAB method's procedural steps, highlighting its utility in overcoming the unique challenges associated with mycobacterial DNA extraction.