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Changes over time for:
Appendix 3


Timeline of Changes
This timeline shows the different versions taken from EUR-Lex before exit day and during the implementation period as well as any subsequent versions created after the implementation period as a result of changes made by UK legislation.
The dates for the EU versions are taken from the document dates on EUR-Lex and may not always coincide with when the changes came into force for the document.
For any versions created after the implementation period as a result of changes made by UK legislation the date will coincide with the earliest date on which the change (e.g an insertion, a repeal or a substitution) that was applied came into force. For further information see our guide to revised legislation on Understanding Legislation.
Status:
EU Directives are published on this site to aid cross referencing from UK legislation. Since IP completion day (31 December 2020 11.00 p.m.) no amendments have been applied to this version.
[Appendix 3 Buffers for test procedures
GENERAL: Unopened sterilized buffers can be stored for up to one year. U.K.
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:
|
| Purpose | Quantity (per L) |
---|
Lubrol flakes | Deflocculant | 0,5 g |
DC silicone antifoam | Anti-foam agent | 1,0 ml |
Tetrasodium pyrophosphate | Anti-oxidant | 1,0 g |
Polyvinylpyrrolidone- 40 000 (PVP-40) | Binding of PCR inhibitors | 50 g |
1.2. Pellet buffer (10 mM phosphate buffer, pH 7,2) U.K.
This buffer is used for resuspension and dilution of potato tuber heel-end core extracts following concentration to a pellet by centrifugation.
Na 2 HPO 4 .12H 2 O | 2,7 g |
NaH 2 PO 4 .2H 2 O | 0,4 g |
Distilled water | 1,00 L |
Dissolve ingredients, check pH and sterilise by autoclaving at 121 °C for 15 min.
2. Buffers for the IF test U.K.
2.1. IF-Buffer (10 mM phosphate buffered saline (PBS), pH 7,2) U.K.
This buffer is used for dilution of antibodies.
Na 2 HPO 4 .12H 2 O | 2,7 g |
NaH 2 PO 4 .2H 2 O | 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 min.
2.2. IF-buffer-Tween U.K.
This buffer is used to wash slides.
Add 0,1 % Tween 20 to the IF buffer.
2.3. Phosphate buffered glycerol, pH 7,6 U.K.
This buffer is used as a mountant fluid on the windows of IF slides to enhance fluorescence.
Na 2 HPO 4 .12H 2 O | 3,2 g |
NaH 2 PO 4 .2H 2 O | 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).]
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