By Nicola Casali, Sabine Ehrt (auth.), Tanya Parish, Neil G. Stoker (eds.)
With one-third of the world's inhabitants contaminated with Mycobacterium tuberculosis, over million humans a 12 months demise from tuberculosis, and the looks of multidrug-resistant traces, the necessity to comprehend the biology of M. tuberculosis, and as a way to boost new interventions, has turn into acute. In Mycobacterium tuberculosis Protocols, best investigators with wide sensible wisdom and adventure describe their most sensible tools for learning this risky pathogen. choked with step by step directions to make sure profitable effects, those equipment variety from uncomplicated dealing with options to the applying of sensible genomics. those molecular strategies are appropriate for learn in genetics, biochemistry, microbiology, telephone biology, epidemiology, and diagnostics, and are on the leading edge of organic examine as an entire, in addition to in targeted M. tuberculosis study. Highlights contain equipment for the elemental defense and tradition of M. tuberculosis, fractionation of the bacterium (nucleic acids, lipids, tradition filtrate, and capsule), the research of gene expression (start-site mapping, real-time PCR, microarrays, and proteomics), the expansion of the bacterium in macrophages and coffee oxygen, cytological research of the micro organism, and diagnostics. hugely sensible and available, Mycobacterium tuberculosis Protocols makes use of complex useful genomics and mutagenesis methodologies to supply either experimental and medical investigators the entire strong strategies had to remove darkness from the molecular biology of tuberculosis and its interactions with host cells, and so force paintings at the wide array of rising healing opportunities.
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Additional info for Mycobacterium tuberculosis Protocols
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Centrifugation of samples containing live M. tuberculosis should be carried out in holders which seal with O-rings to minimize danger of creating aerosol suspensions. 6. Heat treatment at 80°C for 1 h will effectively kill the pathogen (18). Nevertheless, for added safety, subsequent steps (up to the phenol extraction step) should be carried out in a laminar flow cabinet. Extending the period of heating may lead to DNA degradation. 7. Decontamination of the microtome knife can be done with 70% ethanol after sectioning each block.
1998) Identification of differentially expressed mRNA in prokaryotic organisms by customized amplification libraries (DECAL): the effect of isoniazid on gene expression in Mycobacterium tuberculosis. Proc. Natl. Acad. Sci. USA 95, 13,227–13,232. 4. Graham, J. , and Clark-Curtiss, J. E. (1999) Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS). Proc. Natl. Acad. Sci. USA 96, 11,554–11,559.