Abstract
Here, we describe a method for a large-scale liquid screening approach in C. glabrata. This liquid media method offers several distinct advantages over solid media approaches. This includes growth measurement on a plate reader instead of comparing growth by eye-sight. Furthermore, the liquid method requires lower amounts of antifungals and offers a higher sensitivity. While this method has been optimized for C. glabrata it might be used for other Candida species and yeast-like fungi as well.
Keywords: Candida glabrata, Liquid media screening, Antifungals, Large-scale screening, Antifungal tolerance, Antifungal resistance
Materials and Reagents
Equipment
Procedure
Figure 1. Overview of the plates used in the screening process. A. Single colonies are picked with sterile pipette tips and used to inoculate the wells on a 96-well plate filled with liquid YPD medium. After incubation overnight, this plate is used to inoculate several 96-well plates that are later used as stock plates and frozen at -80 °C. B. A stock plate can be thawed and used to inoculate a plate with fresh media. After 24 h of incubation this plate holds the culture that is used to inoculate the plates for the (C) actual screening. Note: All of the working steps shown in this section should be performed under sterile conditions unless stated otherwise.
Notes
Recipes
Acknowledgments
This work was supported by the Austrian Science Foundation FWF through the ERA-Net Pathogenomics project FunPath (FWF-API-0125), and in part by grants from the Christian Doppler Society, the FP7 EC project FUNGITECT, the Marie-Curie ITN ImResFun (MC-ITN-606786) and the FWF Project FWF-P25333 ‘Chromatin’ to KK.
References
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