Council Directive 98/57/EC of 20 July 1998 on the control of Ralstonia solanacearum (Smith) Yabuuchi et al. (c. 57)

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[F1 ANNEX II U.K. TEST SCHEME FOR DIAGNOSIS, DETECTION AND IDENTIFICATION OF RALSTONIA SOLANACEARUM (SMITH) YABUUCHI ET AL.

Textual Amendments

F1 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..

Appendix 6 Validated PCR protocols and reagents

1. PCR protocol of Seal et al. (1993) U.K.

1.2. PCR reaction mix U.K.

a Method was validated using Taq polymerase from Perkin Elmer (AmpliTaq) and Gibco BRL.]
Reagent	Quantity per reaction	Final concentration
Sterile UPW	17,65 µl
10X PCR buffer a (15 mM MgCl 2 )	2,5 µl	1X (1,5 mM MgCl 2 )
dNTP mix (20 mM)	0,25 µl	0,2 mM
Primer OLI-1 (20 µM)	1,25 µl	1µM
Primer Y-2 (20 µM)	1,25 µl	1µM
Taq polymerase (5U/µl) a	0,1 µl	0,5 U
Sample volume	2,0 µl
Total volume	25 µl