Danilchik's for Amy (DFA) Medium (pH 8.3)
| Reagent | Final concentration |
|---|---|
| NaCl | 53 mm |
| Na2CO3 | 5 mm |
| Potassium gluconate | 4.5 mm |
| Sodium gluconate | 32 mm |
| CaCl2 | 1 mm |
| MgSO4 | 1 mm |
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Adjust the pH to 8.3 with granular bicine. Filter with a 0.22 µm filter. Include 0.1% bovine serum albumen (BSA; Sigma-Aldrich A7906) to prevent nonspecific adhesion to substrates or glass coverslips. Just prior to use, add 1% of antibiotic/antimycotic (Sigma-Aldrich A5955). Prepare the DFA with BSA, aliquot, and store at −20°C. Keep aliquots of antibiotic/antimycotic at −20°C and add to thawed DFA before use. Replace DFA with medium containing fresh antibiotic/antimycotic if culturing explants for >24 h. (This recipe is adapted from Sater et al. 1993.)
