By Erika J. Ernst
A suite of state of the art molecular equipment for learning antifungal resistance, for locating and comparing either new and latest antifungal medicinal drugs, and for realizing the host reaction and immunotherapy of such brokers. The protocols stick with the profitable tools in Molecular drugs™ sequence layout, each one supplying step by step laboratory directions, an advent outlining the primary at the back of the strategy, lists of the required gear and reagents, and tips about troubleshooting and heading off identified pitfalls. Antifungal brokers: tools and Protocols deals clinician-scientists, microbiologists and molecular biologists the efficient instruments they want this day to appreciate and effectively advance new healing brokers for yeast, mould, and fungal infections.
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Additional resources for Antifungal Agents: Methods And Protocols (Methods in Molecular Medicine)
San Francisco. 11. Maiden, M. , Bygraves, J. , Russell, J. , et al. (1999) Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc. Natl. Acad. Sci. USA. 95, 3140–3145. 12. Bougnoux, M. , and d’Enfert, C. (2002) Usefulness of multilocus sequence typing for characterization of clinical isolates of Candida albicans. J. Clin. Microbiol. 40, 1290–1297. 13. , and McClelland, M. (1990) Fingerprinting genomes using PCR with arbitrary primers.
Use pRFL37 described by Lasker et al. (3) as a source of the CARE-2 element instead of amplifying it from genomic DNA of C. albicans to make sure that you always use the same sequence as probe. For optimal comparison of hybridization patterns use the ethidium bromide-stained gel to check that equal amounts of DNA have been loaded for all strains. Acknowledgments We thank Katherine Barker for critical reading of the manuscript. Work in our laboratories is supported by the Deutsche Forschungsgemeinschaft, the Bundesministerium für Bildung und Forschung, the Bayerische Forschungsgemeinschaft, the European Union, and the National Institutes of Health.
3. Chloroform. Genomics of Azole Resistance in C. albicans 10. 11. 12. 13. 14. 15. 16. 47 Isopropanol. 0. Diethylpyrocarbonate (DEPC)-treated water. 5 or 2 mL RNase-free microcentrifuge tubes. RNase-free micropipetor tips. Agarose. 2 M MOPS, 100 mM sodium acetate, 10 mM EDTA. 2. Probe Preparation and Microarray Hybridization 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. Deoxynucleotides. T20VN primer. Tris-HCl. Dithiothreitol. Potassium chloride. 1 M Magnesium chloride solution.
Antifungal Agents: Methods And Protocols (Methods in Molecular Medicine) by Erika J. Ernst