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Real-Time PCR (qPCR)
AAT Bioquest offers a comprehensive portfolio of real-time PCR (qPCR) reagents designed for accurate and reproducible nucleic acid quantification. Our product line includes ready-to-use master mixes featuring TAQuest™ formulations for both dye-based and probe-based detection, Helixyte™ Green DNA-binding dyes (equivalent to SYBR® Green I), ROX reference standards for signal normalization, calibration plates and solutions for instrument maintenance, and essential building blocks for custom probe synthesis including phosphoramidites, quenchers, and minor groove binders.
Product Type | Detection Method | Best For | Key Products |
|---|---|---|---|
qPCR Master Mixes (Dye-Based) | Helixyte™ Green | Cost-effective gene expression, melt curve analysis | TAQuest™ Master Mixes with Helixyte™ Green |
qPCR Master Mixes (Probe-Based) | TaqMan® Probes | Multiplexing, high specificity, SNP detection | TAQuest™ Master Mixes for TaqMan Probes |
DNA-Binding Dyes | Minor-groove binding dye | Custom qPCR formulations, gel staining | Helixyte™ Green, Q4ever™ Green |
ROX Reference Dyes | Passive reference | Signal normalization on ABI instruments | 6-ROXtra™, ROX Reference Dye |
Calibration Plates & Solutions | Instrument calibration | ABI 7500 Fast system maintenance | qPCR Calibration Plates (FAM, ROX, VIC, etc.) |
Probe Synthesis Building Blocks | Oligonucleotide synthesis | Custom TaqMan probe manufacturing | Tide Quencher™ CPGs, VIC phosphoramidites |
Viability PCR Reagents | Photo-crosslinking | Live/dead microbial discrimination | MycoLight™ PMA, MycoLight™ vPCR350 |
Fig. 1
Like conventional PCR, qPCR utilizes the same three-step amplification procedure - denaturation, annealing, and extension (figure made in BioRender).
TAQuest™ qPCR Master Mixes
TAQuest™ Master Mixes are optimized 2X ready-to-use solutions containing hot-start Taq DNA polymerase, dNTPs, MgCl₂, and stabilizers for reliable qPCR performance. Available in both standard and FAST cycling formulations, these master mixes are offered with Helixyte™ Green for dye-based detection or optimized for TaqMan probe-based assays. ROX reference dye options (No ROX, Low ROX, High ROX) accommodate different instrument requirements.
Key Features
- Hot-start technology minimizes non-specific amplification and primer-dimer formation
- Room temperature stability allows convenient reaction setup
- High sensitivity enables detection from low template inputs
- Compatible with fast cycling protocols (FAST versions) for reduced run times
- ROX options available for ABI, Bio-Rad, Roche, and other platforms
Fig. 2
Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ FAST qPCR Master Mix with Helixyte™ Green *Low ROX*.
Helixyte™ Green DNA-Binding Dyes
Helixyte™ Green is AAT Bioquest's high-performance DNA-binding dye with the same chemical structure as SYBR® Green I. Upon binding to double-stranded DNA, Helixyte™ Green exhibits dramatic fluorescence enhancement (>1000-fold), enabling sensitive real-time detection of PCR amplicons. Available in multiple concentrations for qPCR applications or as an NHS ester for conjugation to other molecules.
Key Features
- Equivalent to SYBR® Green I with identical spectral properties (Ex/Em = 498/522 nm)
- >1000-fold fluorescence enhancement upon dsDNA binding
- Compatible with all qPCR instruments using SYBR/FAM filter sets
- Cost-effective alternative to SYBR® Green branded products
- Multiple formats available including 20X and 10,000X concentrates
Fig. 3
Quantitative PCR results targeting GAPDH with an input of 100 ng-0.00001 ng cDNA were performed using Helixyte™ Green 20X Aqueous PCR Solution (Catalog Number 17591) and a Fast Advanced Master Mix on an Applied Biosystems® 7500 FAST Real-Time PCR System.
ROX Reference Dyes
ROX (rhodamine X) reference dyes serve as passive internal standards in qPCR reactions to normalize fluorescence signals and correct for well-to-well variations caused by pipetting errors, optical path differences, or other non-PCR-related fluctuations. 6-ROXtra™ is AAT Bioquest's enhanced ROX formulation offering significantly improved stability compared to standard 6-ROX, which is known to degrade over time.
Key Features
- 6-ROXtra™ provides superior stability compared to standard 6-ROX dyes
- Essential for ABI instruments that require ROX normalization
- Pre-diluted and ready-to-use formulations available
- Multiple concentration options (25 µM solutions, 50X concentrates)
Fig. 4
Stability comparison of PCR reference solutions (6-ROX, 6-ROXtra™, and 6-ROX analog). The blue bar represents starting fluorescence. The red bar represents the remaining fluorescence after 4 weeks at 25°C.
qPCR Calibration Plates and Solutions
Real-time PCR instruments require periodic calibration to generate accurate spectral profiles for fluorescence detection. AAT Bioquest provides pre-filled calibration plates and concentrated calibration solutions for the ABI 7500 Fast 96-well system. These products ensure proper dye recognition and accurate multicolor compensation for multiplexed qPCR assays.
Phosphoramidites for Probe Synthesis
Phosphoramidites are essential building blocks for synthesizing fluorescent oligonucleotide probes used in qPCR. VIC phosphoramidites enable the addition of VIC fluorophore to custom oligonucleotides during automated DNA synthesis, while SIMA phosphoramidite provides improved chemical stability for synthesizing HEX-equivalent probes.
Tide Quencher™ Products for FRET Probes
Tide Quencher™ (TQ) molecules are non-fluorescent dark quenchers designed for FRET-based qPCR probes. When paired with appropriate fluorophores, TQ molecules efficiently quench fluorescence until probe hydrolysis during PCR amplification releases the fluorophore. Available as CPGs (controlled pore glass) for direct oligonucleotide synthesis or as reactive forms for post-synthetic conjugation.
Key Features
- Broad quenching range from visible to NIR wavelengths
- TQ5.1 optimal for quenching Cy5, Cy5.5, and similar NIR dyes
- TQ7.1/TQ7.2 designed for far-red fluorophores (Cy7, Alexa Fluor® 750)
- TQ8 optimized for IR fluorophores (ICG, iFluor® 820)
- Multiple reactive forms including CPG, NHS ester, maleimide, and alkyne
Minor Groove Binders (MGB)
Minor Groove Binder (MGB) molecules selectively bind to the minor groove of DNA, increasing the melting temperature (Tm) of oligonucleotide probes. This allows the design of shorter probes with enhanced specificity, particularly useful for SNP detection and allelic discrimination assays. CDPI3 is a tripeptide MGB that can be attached to the 3' end of TaqMan probes.
PCR Reagents and Accessories
AAT Bioquest provides essential reagents for PCR reactions including high-purity dNTPs, reverse transcription kits for cDNA synthesis, PCR reaction buffers, and visualization dyes for tracking reaction mixtures.
This document (01.0101.211015r2) was last updated on Thu Feb 12 2026. All trademarks and registered trademarks mentioned herein are the property of their respective owners.