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Textual Amendments
Basal medium | |
Casamino acids (Difco) | 1,0 g |
Bacto-Peptone (Difco) | 10,0 g |
Glycerol | 5,0 ml |
Bacto-Agar (Difco); see Note 2 . | 15,0 g |
Distilled water | 1,0 L |
Dissolve ingredients and sterilise by autoclaving at 121 °C for 15 minutes.
Cool to 50 °C and add filter-sterilised aqueous stock solutions of the following ingredients to obtain the specified final concentrations:
Crystal Violet (Sigma) | 5 mg per l |
Polymixin-B-Sulphate (Sigma P-1004) | 600 000 U (approximately 100 mg) per l |
Bacitracin (Sigma B-0125) | 1 250 U (approximately 25 mg) per l |
Chloramphenicol (Sigma C-3175) | 5 mg per l |
Penicillin-G (Sigma P-3032) | 825 U (approximately 0,5 mg) per l |
2,3,5-triphenyl tetrazolium chloride (Sigma) | 50 mg per l |
Store media and stock solutions of antibiotics at 4 °C in the dark and use within one month.
Plates should be free from surface condensation before use.
Avoid excess drying of plates.
Quality control should be performed after preparation of each new batch of medium by plating a suspension of a reference culture of R. solanacearum (see Appendix 3) and observing formation of typical colonies after incubation at 28 °C for two to five days.]