{
  "url": "https://openlegislation.uk/eu-directive-2006-56/annex-I/appendix-2",
  "json_url": "https://openlegislation.uk/eu-directive-2006-56/annex-I/appendix-2.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/2",
  "kind": "appendix",
  "number": "2",
  "heading": "Preparation of positive and negative controls for the core screening tests PCR/IF and FISH",
  "text": "Produce a 72 hour culture of a virulent strain of C. m. subsp. sepedonicus (NCPPB 4053 or PD 406) on MTNA basal medium and suspend in 10 mM phosphate buffer to obtain a cell density of approximately 1 to 2 × 108 cfu per ml. This is usually obtained by a faintly turbid suspension equivalent to an optical density of 0,20 at 600 nm.\nRemove the heel end cores of 200 tubers taken from a white skin variety production known to be free from C. m. subsp. sepedonicus.\nProcess the heel ends as usual and resuspend the pellet in 10 ml.\nPrepare 10 sterile 1,5 ml microvials with 900 µl of the resuspended pellet.\nTransfer 100 µl of the suspension of C. m. subsp. sepedonicus to the first microvial. Vortex.\nEstablish decimal levels of contamination by further diluting in the next five microvials.\nThe six contaminated microvials will be used as positive controls. The four non-contaminated microvials will be used as negative controls. Label the microvials accordingly.\nPrepare aliquots of 100 µl in sterile 1,5 ml microvials thus obtaining nine replicas of each control sample. Store at -16 to -24 °C until use.\nThe presence and quantification of C. m. subsp. sepedonicus in the control samples should be first confirmed by IF.\nFor the PCR test perform DNA extraction from positive and negative control samples with each series of test samples.\nFor IF and FISH tests perform assays on positive and negative control samples with each series of test samples.\nFor IF, FISH and PCR assays C. m. subsp. sepedonicus must be detected in at least the 106 and 104 cells/ml of the positive controls and not in any of the negative controls.",
  "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/2",
    "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."
}