{
  "url": "https://openlegislation.uk/eu-directive-1993-85/annex-I/appendix-3",
  "json_url": "https://openlegislation.uk/eu-directive-1993-85/annex-I/appendix-3.json",
  "legislation": "eudr/1993/85",
  "legislation_title": "Council Directive 93/85/EEC of 4 October 1993 on the control of potato ring rot",
  "legislation_url": "https://openlegislation.uk/eu-directive-1993-85",
  "path": "annex/I/appendix/3",
  "kind": "appendix",
  "number": "3",
  "heading": "Buffers for test procedures",
  "text": "GENERAL: Unopened sterilized buffers can be stored for up to one year.\n1.\nBuffers for extraction procedure\n1.1.\nExtraction buffer (50 mM phosphate buffer, pH 7,0)\nThis buffer is used for extraction of the bacterium from plant tissues by homogenisation or shaking.\nNa 2 HPO 4 (anhydrous)\n4,26 g\nKH 2 PO 4\n2,72 g\nDistilled water\n1.00 L\nDissolve ingredients, check pH and sterilise by autoclaving at 121 °C for 15 min.\nAdditional components may be useful as follows:\nFor use with homogenisation extraction method.\nPurpose\nQuantity (per L)\nLubrol flakes\nDeflocculant\n0,5 g\nDC silicone antifoam\nAnti-foam agent\n1,0 ml\nTetrasodium pyrophosphate\nAnti-oxidant\n1,0 g\nPolyvinylpyrrolidone- 40 000 (PVP-40)\nBinding of PCR inhibitors\n50 g\n1.2.\nPellet buffer (10 mM phosphate buffer, pH 7,2)\nThis buffer is used for resuspension and dilution of potato tuber heel-end core extracts following concentration to a pellet by centrifugation.\nNa 2 HPO 4 .12H 2 O\n2,7 g\nNaH 2 PO 4 .2H 2 O\n0,4 g\nDistilled water\n1,00 L\nDissolve ingredients, check pH and sterilise by autoclaving at 121 °C for 15 min.\n2.\nBuffers for the IF test\n2.1.\nIF-Buffer (10 mM phosphate buffered saline (PBS), pH 7,2)\nThis buffer is used for dilution of antibodies.\nNa 2 HPO 4 .12H 2 O\n2,7 g\nNaH 2 PO 4 .2H 2 O\n0,4 g\nNaCl\n8,0 g\nDistilled water\n1.0 L\nDissolve ingredients, check pH and sterilise by autoclaving at 121 °C for 15 min.\n2.2.\nIF-buffer-Tween\nThis buffer is used to wash slides.\nAdd 0,1 % Tween 20 to the IF buffer.\n2.3.\nPhosphate buffered glycerol, pH 7,6\nThis buffer is used as a mountant fluid on the windows of IF slides to enhance fluorescence.\nNa 2 HPO 4 .12H 2 O\n3,2 g\nNaH 2 PO 4 .2H 2 O\n0,15 g\nGlycerol\n50 ml\nDistilled water\n100 ml\nAnti-fading mountant solutions are commercially available e.g. Vectashield ® (Vector Laboratories) or Citifluor ® (Leica).",
  "extent": "E+W+S+N.I.",
  "status": null,
  "version": "current",
  "annotations": [
    {
      "type": "F",
      "label": "F1",
      "text": "Substituted by 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."
    },
    {
      "type": "footnote",
      "label": "f1",
      "text": "For use with homogenisation extraction method."
    }
  ],
  "outstanding_effects": [],
  "provenance": {
    "source": "legislation.gov.uk",
    "source_url": "https://www.legislation.gov.uk/eudr/1993/85/annex/I/appendix/3",
    "retrieved_at": "2026-11-01T15:23:17.858184+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."
}