ANNEX ITEST 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 3Buffers for test procedures

1.Buffers for extraction procedure

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

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

Na2HPO4 (anhydrous)

4,26 g

KH2PO4

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:

Purpose

Quantity (per L)

Lubrol flakes

Deflocculant15

0,5 g

DC silicone antifoam

Anti-foam agent15

1,0 ml

Tetrasodium pyrophosphate

Anti-oxidant

1,0 g

Polyvinylpyrrolidone-40 000 (PVP-40)

Binding of PCR inhibitors

50 g

For use with homogenisation extraction method.