Procedures: PCR Amplification, RFLP and Electrophoresis

 PCR Amplification

The primer pairs used to amplify the ITS (ITS1/ITS4) have been described by White et al. (1990). The oligonucleotide primers used to amplify the IGS were 5SA and CNL12 (Duchesne & Anderson, 1990; see Henrion et al., 1992 for the sequences). Primer CNL12 anneals at the 3' end of the 25S rRNA gene and is extended in the downstream direction whereas primer 5SA anneals at the 5' end of the 5S rDNA. The primers were synthesised and supplied by Bioprobe Systems (Montreuil-sous-Bois, France).

The PCR procedure for in vitro amplification of template DNA has been previously described in detail (Mullis & Faloona, 1987; Saiki et al., 1988).

 RFLP and Electrophoresis