PCR ampli®cation
Each PCR mixture (50 ll) contained 10±25 ng DNA,
200 lM each dNTP (Flowgen), 1 lM each primer, 1X
Dynazyme buffer and 2á0 U Dynazyme (Flowgen). Primer
pairs NS1/NS8 and ITS1/ITS4 (White et al. 1990)
designed for the ampli®cation of speci®c regions of the
rDNA and CHS1/CHS2 (Bowen et al. 1992) for the
conserved regions of chitin synthase genes were used in
PCR. Ampli®cation reactions were performed in a Hybaid
Omnigene thermal cycler (Hybaid, Ashford, Middlesex,
UK). An initial denaturation for 2 min at 95°C was followed
by 35±45 cycles of 1 min at 94°C, 2 min at 55°C and 2 min
at 72°C and a ®nal extension of 5 min at 72°C.