Figure 2.

Example of northern blot hybridisation detecting small RNA (A) and mRNA (B) fractions from wild-type (WT) and a transgenic Gus expressing Fusarium oxysporum lines. For small RNA northern analysis 15 μg of total RNA were separated on a 17% polyacrylamide gel and probed for the U6 small nuclear RNA. For mRNA northern analysis, 10 μg of total RNA were hybridised with a probe detecting sense Gus mRNA. The ethidium staining of the rRNA is shown as loading control. (C) Total RNA from Fusarium oxysporum was used for reverse transcription and PCR to amplify parts of three independent Fusarium genes (FOXG_13826, FOXG_00507, FOXG_09093; Broad Institute Fusarium Comparative Database). Total RNA samples were DNase treated and reverse transcription and PCR carried out using the One-Step RT-PCR Kit (Qiagen, Chadstone, VIC, Australia) using gene specific primers (left panel). A control PCR to confirm that the RNA was free of DNA contamination was performed prior to the reverse transcription reaction (right panel).