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.
Description
Performance Data
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.
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Details
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Contents
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Storage & Handling
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Instrument Capability
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Related Resources
Ordering Information
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PerfeCTa MultiPlex qPCR SuperMix Low ROX50 x 50 μL rxns (1 x 1.25 mL)200 x 50 μL rxns (4 x 1.25 mL)1000 x 50 μL rxns (1 x 25 mL)
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PerfeCTa MultiPlex qPCR SuperMix50 x 50 μL rxns (1 x 1.25 mL)200 x 50 μL rxns (4 x 1.25 mL)1000 x 50 μL rxns (1 x 25 mL)