Council Directive 93/85/EECShow full title

Council Directive 93/85/EEC of 4 October 1993 on the control of potato ring rot

[F1(b) Validated selective growth media U.K.

MTNA medium U.K.

Unless otherwise stated all media components are from BDH.

Yeast extract (Difco) 2,0 g
Mannitol 2,5 g
K 2 HPO 4 0,25 g
KH 2 PO 4 0,25 g
NaCl 0,05 g
MgSO 4 .7H 2 O 0,1 g
MnSO 4 .H 2 O 0,015 g
FeSO 4 .7H 2 O 0,005 g
Agar (Oxoid no. 1) 16,0 g
Distilled water 1,0 L

Dissolve ingredients, adjust pH to 7,2. After autoclaving (at 121 °C for 15 minutes) and cooling down to 50 °C, add the antibiotics: trimethoprim 0,06 g, nalidixic acid 0,002 g, amphotericin B 0,01 g.

Stock antibiotic solutions: trimethoprim (Sigma) and nalidixic acid (Sigma) (both at 5 mg/ml), in 96 % methanol, amphotericin B (Sigma) (1 mg/ml) in dimethyl sulfoxide. Stock solutions are filter-sterilized.

Note: U.K.

Durability of basal medium is three months. After antibiotics are added durability is one month when stored refrigerated.

NCP-88 medium U.K.
Nutrient agar (Difco) 23 g
Yeast extract (Difco) 2 g
D-mannitol 5 g
K 2 HPO 4 2 g
KH 2 PO 4 0,5 g
MgSO 4 .7H 2 O 0,25 g
Distilled water 1,0 L

Dissolve ingredients, adjust pH to 7,2. After autoclaving and cooling down to 50 °C, add the following antibiotics: Polymyxin B sulphate (Sigma) 0,003 g, nalidixic acid (Sigma) 0,008 g, Cycloheximide (Sigma) 0,2 g.

Dissolve antibiotics in stock solutions as follows: nalidixic acid in 0,01 M NaOH, cycloheximide in 50 % ethanol, polymyxin B sulphate in distilled water. Stock solutions are filter-sterilized.

Note: U.K.

Durability of basal medium is three months. After antibiotics are added durability is one month when stored refrigerated.]