Details ofVerticillium 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.

AEach 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.BThe 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.CAssay does not differentiate V. dahliae from V. longisporum lineage A1/D3.