DNA Isolation from Mycobacteria

Methods Mol Biol. 2021:2314:59-75. doi: 10.1007/978-1-0716-1460-0_2.

Abstract

A vast array of molecular biology tools have been developed to investigate the Mycobacterium tuberculosis genome since the advent of its successful sequencing in 1998. These tools, such as quantitative and end point polymerase chain reaction, chromatin immunoprecipitation, and whole genome sequencing, require genomic DNA extracted from lysed mycobacteria. There are numerous methods described in the literature using mechanical, enzymatic, or chemical means to lyse cells and extract genomic DNA to varying degrees of purity. Here, we describe appropriate methods for genomic DNA isolation from solid or liquid cultures from both M. tuberculosis and nontuberculous mycobacteria.

Keywords: Chromatin immunoprecipitation; DNA; Next-generation sequencing; Precipitations; Purification; Quantification; Quantitative PCR; Standards.

MeSH terms

  • Bacteriological Techniques*
  • Cetrimonium / chemistry
  • Chromatin Immunoprecipitation*
  • DNA, Bacterial / isolation & purification*
  • Mycobacterium / genetics*
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA

Substances

  • DNA, Bacterial
  • Cetrimonium