By Richard Calderone, Ronald Cihlar
This quantity offers certain dialogue of various very important concepts that researchers use to check fungal molecular biology and pathogenesis. Written for the Methods in Molecular Biology sequence, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, easily reproducible laboratory protocols, and pointers on troubleshooting and keeping off identified pitfalls.
Authoritative and useful, Candida Species: tools and Protocols aims to make sure winning leads to the extra research of this important field.
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Additional info for Candida Species: Methods and Protocols
Microarrays are particularly well suited for use with this type of impure sample, as C. albicans-specific hybridization probes reduce the contribution from other species. The use of C. albicans-specific primers serves a similar function during RT-qPCR. While RNA-seq lacks this second enrichment step, sequencing reads from other species can be filtered out during analysis by aligning reads to the C. albicans genome. Chemical impurities from the diet and the digestive process may include lipids, bile acids, and other substances that could interfere with downstream applications.
If A and B interacted, this would bring together the two domains of Gal4p, creating a functional transcription factor that switches on a reporter gene. ), Candida Species: Methods and Protocols, Methods in Molecular Biology, vol. 1007/978-1-4939-3052-4_3, © Springer Science+Business Media New York 2016 31 Jaideep Mallick et al. 32 many interacting proteins but had limitations in analyzing proteinprotein interactions among transcription factors, many of which were capable of switching on the transcription of the reporter by themselves without the need of an interacting partner.
11. 67 % SDS. 12. 1 % Triton X-100. 13. 0). 14. 0). 15. Protein-A Sepharose beads suspension: Transfer 1 ml of Protein-A Sepharose beads (shipped as ethanol slurry) into a 15 ml disposable conical tube using a cut-off tip. 0) stored at 4 ºC. 0) and gently rotate on a rotator mixer at 4 °C for 10 min. Spin down beads at 4 °C, 2750 × g, 10 min. Aspirate supernatant. Repeat wash and spin. Aspirate supernatant. 0) to generate 50 % slurry. Store the beads at 4 °C. 3 Equipment 1. Conical flasks. 2. Centrifuge (benchtop).