This document provides an optimized protocol for using the Stratagene Brilliant III Ultra-Fast QRT-PCR Master Mix with the ABI 7500 Fast Real-Time PCR System, and it serves as a quick reference guide for the users.
This document serves as a quick reference guide for the Stratagene Brilliant III Ultra-Fast QRT-PCR Master Mix, now under Agilent Technologies. It details the preparation of reactions, including diluting the reference dye and combining reagent components like the master mix, primers, and RNA templates to a final volume of 20 µl. The guide specifies that reagent mixtures should be kept on ice and mixed gently to avoid bubbles.
| Reaction Volume | 20 µl |
|---|---|
| Total RNA Quantity | 0.1 pg – 100 ng |
| mRNA Quantity | 0.1 pg – 1 ng |
| Reference Dye Dilution | 1:500 |
| Probe Concentration | 100–600 nM |
| Primer Concentration | 200–600 nM |
| Catalog #400 reactions | 600 884 |
| Catalog #4000 reactions | 600 885 |
It is optimized for use with the 7500 Fast Real-Time PCR System from Applied Biosystems.
A new step needs to be added to the beginning of the profile for a 50°C incubation.
The mixture should be gently mixed without creating bubbles and kept on ice.