Fig 5. Real-time PCR amplification of Cronartium spp. spores collected on pines.
Average Ct values obtained with Cronartium spores, contrasting fresh and lyophilized reagents, and field-ready vs. kit DNA extraction. A) Results using spores from C. comandrae. Spores from two different blisters were used, labelled as sample #1 and sample #2. DNA from spores from each sample was extracted using a DNA extraction kit or using the field-ready Edwards buffer. B) Results using spores from C. ribicola. Spores were pooled from multiple blisters. Four extractions were performed and tested, two using a DNA extraction kit and two using field-ready Edwards buffer. C) Results obtained using a portable real-time PCR instrument. Two DNA extractions were prepared in the instrument and tested using lyophilized reagents (extraction #2 and #3). DNA extracted using a kit was used as a positive control (extraction #1). The C. ribicola probe carries the FAM fluorophore and the C. comandrae carries the CY5 fluorophore.