PerfeCTa MultiPlex qPCR SuperMix
Features & Benefits
- Available in two versions: Universal and Low-ROX
- Maximum assay sensitivity and target precision with stringent, ultrapure, and high-yielding AccuStart™ II hot start Taq DNA polymerase
- Broad linear detection range with cDNA (106) or genomic DNA (104) templates
- Enables highly sensitive 4 target detection assays (100) with coamplified, robust internal positive controls (106)
- Supports efficient vortex mixing with proprietary anti-foaming technology
PerfeCTa Multiplex qPCR SuperMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
Faster Cycling with 4-Plex qPCR
Four-color multiplex comparison: fast cycling conditions
Varying amounts of GAPD plasmid DNA (10 to 1 x 107 copies) were co-amplified with 1 x 108 copies (each) of beta-actin (ACTB), interleukin-1beta (IL1B), and α-tubulin (TUB). 4-color multiplexed qPCRs were carried out in 25-µl volumes with PerfeCTa™ Multiplex qPCR SuperMix or QuantiTect® Multiplex PCR Kit (Qiagen). Primer and probe concentrations were according to each manufacturers recommend conditions. Each hot-start Taq was activated according to conditions specified for each product. QuantiTect reagents were incubated for 15 min at 95C, where as PerfeCTa™ Multiplex reactions were incubated for 3 min at 95C. All reactions were cycled for 45 cycles of 10s, 95°C, 30s, 60°C. Average of four replicate reactions for each input amount of GAPD plasmid (FAM-labeled probe) are shown. qPCR of ACTB (HEX-labeled probe), IL1B (Texas Red-Labeled probe), and TUB (Cy5-labelled probe) not shown. All probes utilized non-fluorescent Black Hole Quencher® (BioSearch Technologies). Significant degradation of performance with the QuantiTect reagents using abbreviated cycle times is readily apparent. However, recovery of full and unmodified Taq DNA polymerase activity afforded by PerfeCTa™ Multiplex qPCR SuperMix enables highly efficient multiplex qPCR even with rapid cycling conditions.
4-plex qPCR Comparison
Four-color multiplex comparison: extended cycling conditions.
Varying amounts of GAPD plasmid DNA (10 to 1 x 107 copies) were co-amplified with 1 x 108 copies (each) of beta-actin (ACTB), interleukin-1beta (IL1B), and α-tubulin (TUB). 4-color multiplexed qPCRs were carried out in 50-µl volumes with PerfeCTa™ Multiplex qPCR SuperMix or QuantiTect® Multiplex PCR Kit (Qiagen) according to each manufacturers recommend conditions. Reactions were cycled according to the Qiagen recommended protocol: 15 min, 95°C, followed by 45 cycles of 1 min, 94°C, 1.5 min, 60°C. Average of four replicate reactions for each input amount of GAPD plasmid (FAM-labeled probe) are shown. qPCR of ACTB (HEX-labeled probe), IL1B (Texas Red-Labeled probe), and TUB (Cy5-labelled probe) not shown. All probes utilized non-fluorescent Black Hole Quencher® (BioSearch Technologies). Suppression of GAPD Cts are evident in reactions with the QuantiTect reagents when competing amplicons are >1000-fold excess over the limiting target sequence (GAPD). PerfeCTa™ Multiplex qPCR SuperMix accurately amplifies GAPD target in the presences of >1 x 107-fold excess of competing amplicons.
A PCR Assay Detects Legionella pneumophila Harboring Mobile Element ICE-βox in a Variety of Water Sources
2X concentrated SuperMix Reagent, containing:
- Reaction buffer with multiplex qPCR-optimized MgCl2, dATP, dCTP, dGTP, and dTTP.
- Ultra-pure AccuStart hot start Taq DNA Polymerase.
- Reference dye (if applicable).
- Proprietary performance enhancing additives and stabilizers.
Storage & HandlingStore components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt. Repeated freezing and thawing of the Supermix is not recommended. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
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