Fig 2.

PCR detection of A. fumigatus. (A) PCR amplicons of the aclF1-5′ and aclF1-3′ conserved regions were analyzed by 2% agarose gel electrophoresis. Total genomic DNA templates were used from 12 A. fumigatus strains and 17 non-A. fumigatus controls; strain abbreviations are shown in Table 1. M, 1-kb Plus TrackIt DNA ladder. (B) Two-plex PCR of the aclF1-5′ and -3′ regions. Results of amplification with the A. fumigatus Af293 strain are shown in the first lane. In addition, control genomic DNA from the non-A. fumigatus species was used to demonstrate specificity of two-plex PCR. Genomic DNA templates were standardized to approximately 30 ng/μl. PCR amplification was resolved by 2% agarose gel electrophoresis. M, 1-kb Plus TrackIt DNA ladder. (C) Nanogel electrophoretic separations of two-plex PCR. The DNA base ladder (top trace) and PCR-amplified markers of A. fumigatus (bottom trace) detected with the intercalating dye SYBR green 1 are shown. The separation is accomplished using a 25-μm-inner-diameter capillary with an effective length of 40.2 cm, E = 100 V/cm, 30°C, and a 10% nanogel with [DMPC]/[DHPC] = 2.5 at 6 kV for a 2-s injection.