F1ANNEX IITEST SCHEME FOR DIAGNOSIS, DETECTION AND IDENTIFICATION OF RALSTONIA SOLANACEARUM (SMITH) YABUUCHI ET AL.
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Amendments (Textual)
Appendix 4Buffers for test procedures
2.Buffers for the IF test
2.1.IF-Buffer (10 mM phosphate buffered saline (PBS), pH 7.2)
This buffer is used for dilution of antibodies
Na2HPO4.12H2O | 2,7 g |
NaH2PO4.2H2O | 0,4 g |
NaCL | 8,0 g |
Distilled water | 1,0 l |
Dissolve ingredients, check pH and sterilise by autoclaving at 121 °C for 15 minutes.
2.2.IF-buffer-Tween
This buffer is used to wash slides.
Add 0,1 % Tween 20 to the IF buffer.
2.3.Phosphate buffered glycerol, pH 7,6
This buffer is used as a mountant fluid on the windows of IF slides to enhance fluorescence.
Na2HPO4.12H2O | 3,2 g |
NaH2PO4.2H2O | 0,15 g |
Glycerol | 50 ml |
Distilled water | 100 ml |
Anti-fading mountant solutions are commercially available e.g. Vectashield® (Vector Laboratories) or Citifluor® (Leica).
Substituted by Commission Directive 2006/63/CE of 14 July 2006 amending Annexes II to VII to Council Directive 98/57/EC on the control of Ralstonia solanacearum (Smith) Yabuuchi et al..