By Erika J. Ernst
A suite of state of the art molecular tools for learning antifungal resistance, for locating and comparing either new and present antifungal medicines, and for figuring out the host reaction and immunotherapy of such brokers. The protocols stick to the winning equipment in Molecular drugs™ sequence layout, each one providing step by step laboratory directions, an advent outlining the primary at the back of the strategy, lists of the mandatory gear and reagents, and tips about troubleshooting and averting identified pitfalls. Antifungal brokers: equipment and Protocols deals clinician-scientists, microbiologists and molecular biologists the effective instruments they want this day to appreciate and effectively enhance new healing brokers for yeast, mildew, and fungal infections.
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Extra resources for Antifungal Agents: Methods And Protocols (Methods in Molecular Medicine)
The MPA-sensitive wild-type parental strain was included as a negative control. 5 µg/mL MPA. Targeted Gene Deletion in C. 2. Transformation of C. albicans by Electroporation 1. Prepare an overnight culture of your C. albicans host strain in YPD medium. 2. Inoculate an Erlenmeyer flask containing 50 mL of YPD medium with 5 µL of the preculture and grow overnight at 30°C on a rotary shaker set to 250 rpm. 3. 2, transfer the cells into a sterile 50-mL tube and centrifuge for 5 min at 3300g. 4. Carefully remove the supernatant using a 10-mL glass pipet and resuspend the cells in 8 mL of sterile double-distilled H2O.
Microbiol. 34, 3063–3071. DNA Fingerprinting Methods for Candida Species 25 6. Lockhart, S. , Pfaller, M. , and Soll, D. R. (1997) Development and verification of fingerprinting probes for Candida glabrata. Microbiology 243, 3733–3746. 7. , and Soll, D. R. (1999) Development and characterization of complex DNA fingerprinting probes for the infectious yeast Candida dubliniensis. J. Clin. Microbiol. 37, 1035–1044. 8. , and Soll, D. R. (2001) Cloning and characterization of a complex DNA fingerprinting probe for Candida parapsilosis.
6. 7. 8. Oligo(dT) primer or random hexamer primers. RNase block ribonuclease inhibitor. Moloney murine leukemia virus reverse transcriptase. SuperScript™ First-Stand Synthesis System for RT-PCR (containing RT reaction buffer). Heat block with temperature range between 37 and 100°C. PCR primers. Taq polymerase. 1% Triton X-100 solution). 48 Barker and Rogers 3. Methods DNA microarray analysis allows for the measurement of steady-state messenger ribonucleic acid (mRNA) levels for most genes in the C.