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 (c. 56)

ANNEX I U.K. TEST SCHEME FOR DIAGNOSIS, DETECTION AND IDENTIFICATION OF THE RING ROT BACTERIUM, CLAVIBACTER MICHIGANENSIS (Smith) Davis et al. ssp. SEPEDONICUS (Spieckermann et Kotthoff) Davis et al. SCOPE OF THE TEST SCHEME

6. PCR TEST U.K.

6.2. PCR U.K.

6.2.1. Prepare test and control templates for PCR according to the validated protocol (Appendix 6). Prepare one decimal dilution of sample DNA extract (1:10 in UPW). U.K.

6.2.2. Prepare the appropriate PCR reaction mix in a contamination-free environment according to the published protocol (Appendix 6). The validated PCR protocol is a multiplex reaction that also incorporates an internal PCR control. U.K.

6.2.3. Add 5 µl of DNA extract per 25 µl PCR reaction in sterile PCR tubes. U.K.

6.2.4. Incorporate a negative control sample containing only PCR reaction mix and add the same source of UPW as used in the PCR mix in place of sample. U.K.

6.2.5. Place tubes in the same thermal cycler which was used in preliminary testing and run the appropriately optimised PCR programme (Appendix 6). U.K.