This quick reference guide provides an optimized protocol for using the Stratagene Brilliant III Ultra- Fast SYBR® Green QPCR Master Mix with the Stratagene Mx3000P and Mx3005P QPCR Systems from Agilent.
This document serves as a quick reference guide for the Stratagene Brilliant III Ultra-Fast SYBR Green QPCR Master Mix, designed for use with the Stratagene Mx3000P and Mx3005P QPCR Systems from Agilent Technologies. It provides optimized protocols for preparing reactions, setting up the QPCR plate, and running the PCR program.
The guide details the steps for diluting the reference dye and combining reagent components, including specific quantities for genomic DNA and cDNA. It also outlines the software setup for the thermal profile, specifying settings for the Pre-Melt/RT, Amplification, and Dissociation segments, and provides instructions for running the program and analyzing data.
| Reference Dye Dilution | 1:500 |
|---|---|
| Final Reaction Volume | 20 µl |
| Primer Concentration | 200–500 nM |
| Genomic DNA Quantity | 5 pg – 50 ng |
| cDNA Quantity | 0.5 pg – 100 ng |
| Denaturation Step | 5 seconds |
| Product Catalog # (400 reactions) | 600882 |
| Product Catalog # (4000 reactions) | 600883 |
The reference dye should be diluted 1:500 using nuclease-free PCR-grade water.
The recommended final reaction volume is 20 µl.
The suggested thermal profile includes a 5-second denaturation step.