[F1ANNEX 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

Appendix 3 Buffers for test procedures

1. Buffers for extraction procedure U.K.

1.1. Extraction buffer (50 mM phosphate buffer, pH 7,0) U.K.

This buffer is used for extraction of the bacterium from plant tissues by homogenisation or shaking.

Na 2 HPO 4 (anhydrous) 4,26 g
KH 2 PO 4 2,72 g
Distilled water 1.00 L

Dissolve ingredients, check pH and sterilise by autoclaving at 121 °C for 15 min.

Additional components may be useful as follows:

a

For use with homogenisation extraction method.]

Purpose Quantity (per L)
Lubrol flakes Deflocculant a 0,5 g
DC silicone antifoam Anti-foam agent a 1,0 ml
Tetrasodium pyrophosphate Anti-oxidant 1,0 g
Polyvinylpyrrolidone- 40 000 (PVP-40) Binding of PCR inhibitors 50 g