This quick reference guide provides an optimized protocol for using the Stratagene Brilliant III Ultra-Fast SYBR® Green QRT-PCR Master Mix with the 7500 Fast Real-Time PCR System from Applied Biosystems. For detailed instructions, refer to the full product manual.
The document serves as a quick reference guide for preparing reactions and setting up experiments using the Stratagene Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix. It details steps for diluting the reference dye, combining reagent components including primers and DTT, and adding experimental RNA to achieve a final reaction volume of 20 μl.
| Final reaction volume | 20 μl |
|---|---|
| Upstream primer concentration range | 150–500 nM |
| Downstream primer concentration range | 150–500 nM |
| Diluted reference dye volume per reaction | 0.3 μl |
| DTT concentration in mixture | 100 mM |
The suggested quantity range for Total RNA is 0.1 pg – 100 ng.
The suggested quantity range for mRNA is 0.1 pg – 1 ng.
The reagent mixture should be kept on ice after preparation and mixing.