This fully updated edition explores the latest techniques to study the challenging, and at times dangerous, genus of bacteria known as mycobacteria with basic methods that are still required for mycobacteriology along with the newer or improved methods that have been developed. The volume features chapters on the basics of DNA isolation, protein isolation, and lipid isolation, as well as more sophisticated techniques for isolation of ribosomes, and continues with sections involving analyzing subcellular fractions, culture methods, sequencing technology, in vitro models, molecular methods, as well as drug discovery applications. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Mycobacteria Protocols, Fourth Edition serves as an ideal guide for those starting out with their mycobacteria research and also for those who have worked with it for decades.
Inhaltsverzeichnis
Culturing Mycobacteria. - DNA Isolation from Mycobacteria. - Extraction and Separation of Mycobacterial Proteins. - Lipid and Lipoarabinomannan Isolation and Characterization. - Purification of Hibernating and Active C- Ribosomes from Zinc-Starved Mycobacteria. - Macrophage Infection Models for
Mycobacterium tuberculosis
. - Novel Single-Cell and High Throughput Microscopy Techniques to Monitor
Dictyostelium discoideum
-
Mycobacterium marinum
Infection Dynamics. - Single-Cell Analysis of Mycobacteria Using Microfluidics and Time-Lapse Microscopy. - Measuring Efflux and Permeability in Mycobacteria. - Drug Susceptibility Screening Using In Vitro Models of Hypoxic Non-Replicating Persistent Mycobacteria. - Flow Cytometry Analysis of Mycobacteria and Mycobacteria-Infected Immune Cells. - Electroporation of Mycobacteria. - Gene Switching and Essentiality Testing. - Oligo-Mediated Recombineering and Its Use for Making SNPs, Knockouts, Insertions, and Fusions in
Mycobacterium tuberculosis
. - Using Proteolytic Hypomorphs to Detect Small Molecule Mechanism of Action. - CRISPR Interference (CRISPRi) for Targeted Gene Silencing in Mycobacteria. - Exploiting Fluorescent Proteins to Understand
Mycobacterium tuberculosis
Biology. - Metabolic Labeling of Live Mycobacteria with Trehalose-Based Probes. - Identification and Characterization of Mycobacterial Species Using Whole Genome Sequences. - Whole Genome Sequencing of
Mycobacterium tuberculosis
Directly from Sputum Samples. - Experimental and Computational Workflow for RNA Sequencing in
Mycobacterium tuberculosis
: From Total RNA to Differentially Expressed Genes. - RNA Sequencing for Transcript 5 -End Mapping in Mycobacteria. - Methods for Proteomic Analyses of Mycobacteria. - Targeted Lipidomics of Mycobacterial Lipids and Glycolipids. - Metabolomics of
Mycobacterium tuberculosis
. - Determining Minimum Inhibitory Concentrations in Liquid Cultures or on Solid Medium. - A Multi-Stress Model for High Throughput Screening Against Non-Replicating
Mycobacterium tuberculosis
. - Antimicrobial Susceptibility Testing for
Mycobacterium sps
in High Throughput Format. - High-Content Analysis Monitoring Intracellular Trafficking and Replication of
Mycobacterium tuberculosis
Inside Host Cells. - Efficient Measurement of Drug Interactions with DiaMOND (Diagonal Measurement of N-Way Drug Interactions). - Drug Sensitivity Testing of
Mycobacterium tuberculosis
Growing in a Hollow Fiber Bioreactor.
