Home -> qPCR: TaqMan® Probe Detection
TaqMan® Probe Detection
PerfeCTa™ qPCR SuperMixes are ready-to-use 2x reaction mixes that contain all components, except probe, primer and template for real-time PCR. PerfeCTa™ qPCR SuperMixes are perfected for exceptional sensitivity, reliability and reproducibility through the use of unique and proprietary polymerases, buffers and additives. PerfeCTa™ qPCR SuperMixes are available and optimized for all real-time PCR instrument platforms including those requiring normalization with ROX reference dye. See our Selection Guide for TaqMan Probe Detection or go directly to the corresponding qPCR product list.
- Extreme sensitivity - reliably detect targets of varying abundance over a broad dynamic range
- High fluorescent signal- optimized for TaqMan® and TaqMan® MGB™ probes
- High specificity - AccuStart™ Taq DNA polymerase provides a superior, antibody-mediated, hot-start
- Decontamination option - available with UNG (uracil-N-glycosylase) for elimination of previously-amplified fragments
PerfeCTa™ qPCR SuperMix: Performance Data
PerfeCTa™ qPCR SuperMix™ comparison to Taqman® Gene Expression MasterMix (Applied BioSystems), RPLPO
RPLPO was amplified from log-fold serial dilutions of qScript™-synthesized cDNA from HeLa cell total RNA (100 ng to 100 fg ) with either PerfeCTa™ qPCR SuperMix or TaqMan Gene Expression Master Mix (Applied Biosystems) using the RPLPO TaqMan Endogenous Control primer/probe set (Applied Biosystems) according to each manufacturers protocol. Triplicate plots for each input quantity are shown. PerfeCTa™ qPCR SuperMix provided superior signal and detected equal input target quantities at earlier Cts for more robust and sensitive qPCR with TaqMan MGB™ probe detection chemistry.
More Performance Data
- PerfeCTa™ qPCR SuperMix™ comparison to Taqman® Gene Expression MasterMix (Applied BioSystems), b2-microglobin
- PerfeCTa™ qPCR SuperMix™ comparison to Taqman® Universal MasterMix (Applied BioSystems), b2-microglobin
- PerfeCTa™ qPCR SuperMix™ comparison to Platinum® qPCR SuperMix (Invitrogen), b2-microglobin
- PerfeCTa™ qPCR SuperMix™ comparison to FastStart™ Taqman® Probe Master (Roche), b2-microglobin
- PerfeCTa™ qPCR SuperMix™ comparison to Taqman® Gene Expression MasterMix (Applied BioSystems), ADAR
- PerfeCTa™ qPCR SuperMix™ comparison to Taqman® Universal MasterMix (Applied BioSystems), ADAR
- PerfeCTa™ qPCR SuperMix™ comparison to Platinum® qPCR SuperMix (Invitrogen), ADAR
- PerfeCTa™ qPCR SuperMix™ comparison to FastStart™ Taqman® Probe Master (Roche), ADAR
PerfeCTa™ qPCR SuperMix, UNG: Performance Data
qPCR of single copy gene, genomic DNA template
Comparable amplification efficiency and sensitivity was obtained for amplification of IL1-β from log fold serial dilutions of human genomic DNA (500 ng to 50 pg) using either PerfeCTa™ qPCR SuperMix, UNG or PerfeCTa™ qPCR SuperMix. Reactions contained 300 nM each primer and 150 nM 5'-FAM/3'-BHQ-1 labeled probe (Biosearch Technologies). Four replicate reactions were run for each input quantity with each SuperMix. Reactions were incubated 5 min at 45°C followed by 5 min at 95C to fully denature and partially fragment the gDNA template. PCR cycling was for 40 cycles of 95°C, 15s; 60°C, 1 min.
More Performance Data
PerfeCTa™ MultiPlex qPCR SuperMix: Performance Data
High performance Four-color multiplexed qPCR with wide linear dynamic range and sensitivity
Varying amounts of GAPD plasmid DNA (10 to 1 x 107 copies) were co-amplified with 1 x 108 copies (each) of β-actin (ACTB), interleukin-1β (IL1B), and α-tubulin (TUB). 4-color multiplexed qPCRs were carried out in 50-µl volumes with PerfeCTa™ Multiplex qPCR SuperMix, 150 nM each probe and 300 nM each primer on an iCycler iQ® (Bio-Rad Laboratories). Reactions were incubated at 95°C for 3 min, followed 45 cycles of 95°C, 10s; 58°C, 1 min. GAPD (FAM-labeled probe) results shown in top panel. Linearity and sensitivity of qPCR was maintained with as few as 10 copies of GAPD with no apparent suppression of PCR due to co-amplification of 1 x 107-fold excess amounts of each of the three other amplicons. qPCR of ACTB (HEX-labeled probe), IL1B (Texas Red-Labeled probe), and TUB (Cy5-labelled probe) are shown in each respective panel. All probes utilized non-fluorescent Black Hole Quencher® (BioSearch Technologies).
More Performance Data
- Overcome challenges of target bias in multiplex qPCR. Four-color multiplexed qPCR with comparable performance to single-plex qPCR.
- Four-color multiplex comparison: extended cycling conditions.
- Four-color multiplex comparison: fast cycling conditions
PerfeCTa™ qPCR FastMix™: Performance Data
Fast-cycle TaqMan® qPCR with equal performance to conventional protocols.
RPLPO was amplified from log-fold serial dilutions of qScript™-synthesized cDNA from HeLa cell total RNA (100 ng to 100 fg ) with either PerfeCTa™ qPCR FastMix (fast cycling protocol) or PerfeCTa™ qPCR SuperMix (conventional cycling conditions) and the RPLPO TaqMan Endogenous Control primer/probe set (Applied Biosystems). Average plots for triplicate reactions of each input quantity are shown.
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