Details of<i>Verticillium</i> simplex PCR assays, including target loci, primer pairs, DNA template concentrations, PCR annealing temperatures, numbers of PCR cycles, PCR product sizes, and agarose gel concentrations for gel electrophoresis.

A<p>Each 25 µl PCR reaction contained the following: 1.25 µl of each primer from 10 µM stocks, 12.5 µl Promega master mix, and 10 µl template containing 1, 10 or 100 ng DNA.</p>B<p>The PCR program consisted of a 2 min initial denaturation step at 94°C, 32 or 35 cycles of 10 sec at 94°C, 20 sec at the PCR assay-dependent annealing temperature, and 1 min at 72°C, followed by a final extension of 7 min at 72°C. PCR reactions were set up at room temperature under sterile conditions and run immediately, or were stored in a freezer.</p>C<p>Assay does not differentiate <i>V. dahliae</i> from <i>V. longisporum</i> lineage A1/D3.