Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs

Abstract

Summary: We describe steps for gDNA isolation from mycobacterium strains isolated from guinea pig lungs. We detail steps for infection of guinea pigs with Mycobacterium tuberculosis, followed by in vitro growth, gDNA isolation, and whole genome sequencing. We also describe an ex vivo competition experiment to determine the selective advantage of one strain over another. We include details for WGS and mutation spectrum analysis. The protocol can be used to identify mutations that arise in other pathogenic bacteria.For complete details on the use and execution of this protocol, please refer to Naz et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Genomics
• Sequencing
• Immunology
• Microbiology
• Model organisms