This application note provides detailed instructions on how to use the Stratagene Mx3000P QPCR system to detect and quantify various agricultural and economically important fungi.
This document details the development and testing of a multiplex real-time PCR assay designed to detect and quantify three major genera of mycotoxigenic fungi: Aspergillus, Fusarium, and Penicillium. The assay utilizes genus-specific probes and fluorophores compatible with the Stratagene Mx3000P QPCR system to analyze DNA from stored grain and other agricultural commodities.
The study outlines the design of primers and probes targeting the internal transcribed spacer (ITS) regions of ribosomal DNA. Results demonstrate the assay's ability to detect DNA concentrations ranging from 10 pg to 10 ng, with specific standard curves generated for each fungal genus. A case study illustrates the application of this method in analyzing distiller's grain contaminated with mold.
| Detection Range | 10 pg to 10 ng of DNA |
|---|---|
| Reaction Volume | 20 ul |
| Primer Concentration | 10 pmol |
| Probe Concentration | 5 pmol |
| Annealing Temperature | 70°C |
The assay is designed to detect and quantify Aspergillus, Fusarium, and Penicillium.
The linear detection range is between 10 pg and 10 ng of DNA.
No, this item is intended for Research Use Only and not for use in diagnostic procedures.