This quick reference guide provides an optimized protocol for using the Stratagene Brilliant III Ultra-Fast QRT-PCR Master Mix with the 7500 Fast Real-Time PCR System from Applied Biosystems.
This document is a Quick Reference Guide for the Agilent Technologies (formerly Stratagene) Brilliant III Ultra-Fast QRT-PCR Master Mix. It provides an optimized protocol specifically designed for use with the Applied Biosystems 7500 Fast Real-Time PCR System.
The guide details the preparation of reactions, including diluting the reference dye and combining reagent components such as the master mix, primers, probe, DTT, and RNA template to a final volume of 20 μl. It also outlines the setup steps for the ABI 7500 software, including selecting TaqMan Reagents and Fast ramp speed, configuring the thermal profile with a specific 50°C incubation step, and running the PCR program.
| Final Reaction Volume | 20 μl |
|---|---|
| Reference Dye Dilution | 1:500 |
| Probe Concentration Range | 100–600 nM |
| Primer Concentration Range | 200–600 nM |
| DTT Concentration | 100 mM |
This guide provides an optimized protocol for using the Stratagene Brilliant III Ultra-Fast QRT-PCR Master Mix with the 7500 Fast Real-Time PCR System from Applied Biosystems.
The reference dye must be diluted 1:500 using nuclease-free PCR-grade water before adding to the reaction mixture.