{
  "url": "https://openlegislation.uk/eu-directive-2006-56/annex-I/appendix-5",
  "json_url": "https://openlegislation.uk/eu-directive-2006-56/annex-I/appendix-5.json",
  "legislation": "eudr/2006/56",
  "legislation_title": "Commission Directive 2006/56/EC of 12 June 2006 amending the Annexes to Council Directive 93/85/EEC on the control of potato ring rot",
  "legislation_url": "https://openlegislation.uk/eu-directive-2006-56",
  "path": "annex/I/appendix/5",
  "kind": "appendix",
  "number": "5",
  "heading": "Media for isolation and culture of C. m. subsp. sepedonicus",
  "text": "(a)\nGeneral growth media\nNutrient agar (NA)\nNutrient Agar (Difco)\n23,0 g\nDistilled water\n1,0 L\nDissolve ingredients and sterilise by autoclaving at 121 °C for 15 min.\nNutrient dextrose agar (NDA)\nDifco bacto nutrient agar containing 1 % D(+) glucose (monohydrate). Sterilize by autoclaving at 115 °C for 20 minutes.\nYeast peptone glucose agar (YPGA)\nYeast Extract (Difco)\n5,0 g\nBacto-Peptone (Difco)\n5,0 g\nD(+) Glucose (monohydrate)\n10,0 g\nBacto-Agar (Difco)\n15,0 g\nDistilled water\n1,0 L\nDissolve ingredients and sterilise by autoclaving at 121 °C for 15 min.\nYeast extract mineral salts medium (YGM)\nBacto-Yeast-Extract (Difco)\n2,0 g\nD(+) Glucose (monohydrate)\n2,5 g\nK2HPO4\n0,25 g\nKH2PO4\n0,25 g\nMgSO4.7H2O\n0,1 g\nMnSO4.H2O\n0,015 g\nNaCl\n0,05 g\nFeSO4.7H2O\n0,005 g\nBacto-Agar (Difco)\n18 g\nDistilled water\n1,0 L\nDissolve ingredients and sterilize 0,5 litre volumes of medium by autoclaving at 115 °C for 20 minutes.\n(b)\nValidated selective growth media\nMTNA medium\nUnless otherwise stated all media components are from BDH.\nYeast extract (Difco)\n2,0 g\nMannitol\n2,5 g\nK2HPO4\n0,25 g\nKH2PO4\n0,25 g\nNaCl\n0,05 g\nMgSO4.7H2O\n0,1 g\nMnSO4.H2O\n0,015 g\nFeSO4.7H2O\n0,005 g\nAgar (Oxoid no. 1)\n16,0 g\nDistilled water\n1,0 L\nDissolve 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.\nStock 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.\nNote:\nDurability of basal medium is three months. After antibiotics are added durability is one month when stored refrigerated.\nNCP-88 medium\nNutrient agar (Difco)\n23 g\nYeast extract (Difco)\n2 g\nD-mannitol\n5 g\nK2HPO4\n2 g\nKH2PO4\n0,5 g\nMgSO4.7H2O\n0,25 g\nDistilled water\n1,0 L\nDissolve 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.\nDissolve 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.\nNote:\nDurability of basal medium is three months. After antibiotics are added durability is one month when stored refrigerated.",
  "extent": "E+W+S+N.I.",
  "status": null,
  "version": "current",
  "annotations": [],
  "outstanding_effects": [],
  "provenance": {
    "source": "legislation.gov.uk",
    "source_url": "https://www.legislation.gov.uk/eudr/2006/56/annex/I/appendix/5",
    "retrieved_at": "2026-11-01T15:22:35.204179+00:00"
  },
  "licence": "Open Government Licence v3.0",
  "licence_url": "https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/",
  "attribution": "Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives."
}