A new detection kit can expedite your qPCR process.
If you are testing for Aspergillus in cannabis and other agricultural products, you can obtain rapid and accurate test results with high specificity by using the new Spex CertiPrep® 2-Color Aspergillus Detection Kit for qPCR. The kit is designed to detect the presence of the four most common pathogenic strains of Aspergillus DNA: niger, terreus, flavus, and fumigatus DNA in cannabis species flower, stem and/or leaf tissue(s).
What’s in the kit?
Components of the kit include 2 x Aspergillus detection Master Mix, Aspergillus positive control, internal control, yellow dye (20 x), DNase/RNase/Proteinase free water and ROX. For machines that lack a HEX filter, VIC can be interchangeably used.
How it works
The internal control ensures the sample is free of PCR inhibitors that would lead to false negative results. To help distinguish the Master Mix from the template DNA and from the final product during sample preparation, an inert yellow dye is provided to label the template prior to adding to the Master Mix which contains an inert blue dye. When mixed the resulting solution is green, confirming the presence of all components. The kit works with any qPCR system that supports the two-color channels. It is very sensitive and can detect down to 10 copies of Aspergillus DNA.
Protocol
Once all components in the kit are thawed in ice, the tubes are mixed by inversion to ensure homogeneity. Before first use, ROX is added to the 2X Aspergillus Detection Master Mix to ensure qPCR machine compatibility. Afterwards, 10 μL of the 2X Aspergillus Detection Master Mix is added to designated wells of a qPCR plate. Then, in a separate 1.5 mL tube the following components are mixed:
| Component | Volume (10 µL) |
| 20 X Yellow Dye | 1 µL |
| DNA Template* | 1–6 µL |
| Internal Control Template | 2 µL |
| Water | up to 10 µL |
- For positive control replace DNA template with 2 μL of Aspergillus Positive Control Template. For negative control, do not add any DNA template.
Lastly, after a brief centrifugation, the components are pipetted to wells containing the 2X Aspergillus Detection Master Mix and qPCR is performed using the following conditions:
| Step | Temperature (ºC) | Time | Number of Cycles |
| Denature | 95 | 1 min | 1 |
| qPCR Detection | 95 | 10 sec | 35 |
| 60 | 30 sec |
Results
The results from the qPCR should be available within 55 minutes, depending on the qPCR machine being used.
See the results graphs. Read the complete technical note.
Get your free 2-Color Aspergillus qPCR Detection sample kit available at spex.com.
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