Minimum Inhibitory Concentration (MIC) Assay for Antifungal Drugs

Materials and Reagents

1. Strain to be analyzed
2. Overnight cultures of the strains to be analyzed and parental controls
3. Glucose
   
4. MOPS
5. Yeast extract
   
6. Bactopeptone
7. Yeast nitrogen base
   
8. Media
   1. RPMI medium 1640 (Life Technologies, Gibco^®, catalog number: 31800-089 ) (see Recipes)
      Note: It is the standard medium recommended by the CLSI standard protocol.
      
   2. YPD (see Recipes)
      Note: It is a rich medium commonly used to assess susceptibility of diverse mutants.
      
   3. Synthetic defined medium (see Recipes)
      Note: Supplemented with required amino acids to enable growth of auxotrophic strains is commonly used for experiments requiring plasmid selection.
      

Equipment

1. 96-well tissue culture plate (flat bottom with lid) (SARSTEDT AG, catalog number: 83.1835 )
2. Microplate reader
3. Multichannel pipette [optional: Multichannel electronic pipette (Rainin, model: EDP3-Plus )]
   

Software

1. Soft Max Pro or equivalent spectrophotometer software (Microplate Data Processing)
2. Microsoft Excel (Data Analysis)
3. JavaTree View (Heat Map Generation, http://jtreeview.sourceforge.net/)

Procedure