PRODUCT

Recombinase Polymerase Amplification (RPA) Real-Time Kit

Real-time fluorescence-enabled RPA (exo probes)

2× lyophilized master mix supports real‑time detection via exo probes at 37–42 °C with minimal hands‑on time. Ideal for rapid DNA target quantification and workflow integration with standard instruments.

Exo‑probe real‑time fluorescence
Minimal hands‑on time for reaction at 37–42 °C
Lyophilized 2× mix for easy workflow integration
Ideal for rapid DNA target quantification

Select a Size

Recombinase Polymerase Amplification (RPA) Real-Time Kit (8 rxn)

#MM32RPAEXO-Sm

Specifications Description Quality Resources

Product Number	MM32RPAEXO
Function	RPA with exonuclease and fluorescent probes
For Use In	Real-Time DNA target quantification
Source	Recombinant E. coli
Shipping Conditions	Ambient
Storage Temperature	Ambient
Stability	≥12 months
Delivery Format	Tubes
Intended Use	This product is for research use only

Real-Time DNA Amplification with Fluorescence Monitoring

The RPA Real-Time Kit is a ready-to-use, high-performance solution for ultra-rapid, real-time DNA amplification under isothermal conditions. Operating at constant low temperatures (37°C–42°C), this kit eliminates the need for thermal cycling, enabling sensitive and specific DNA detection in as little as 10-20 minutes. Its lyophilized master mix format ensures stability for ambient transport and storage, making it ideal for point-of-care and field-based applications.

Optimized for fluorescence-based detection, the kit allows real-time monitoring of amplification with strong signal clarity and minimal hands-on time. Its robust formulation ensures efficient target recognition and consistent performance, even with complex or unprocessed sample types, making it a reliable choice for time-critical diagnostics and research applications.

Key Features

Ready-to-Use Lyophilized Master Mixes: Stable for ambient storage and transport, eliminating the need for refrigeration. Perfect for point-of-care and remote settings.
Ultra-Rapid Amplification: Achieves highly sensitive DNA detection in just 10–20 minutes.
Real-Time Monitoring: Optimized for fluorescence-based detection with strong signal clarity.
Robust Performance: High efficiency and specificity across diverse sample types, including crude extracts.
Isothermal Operation: Functions at constant low temperatures (37°C–42°C), removing the need for thermal cycling.
Customizable Components: Tailor high-purity reagents to your unique assay conditions.

Applications

Pathogen Detection: Rapid and reliable amplification of low-copy-number DNA for clinical diagnostics.
Environmental Testing: Ideal for monitoring contaminants using portable devices in field-based settings.
Assay Development: Scalable nucleic acid detection from research to high-throughput workflows.
Flexible Readouts: Compatible with real-time fluorescence, lateral flow, and other detection methods.

Custom Component Options

We offer flexible customization to meet your specific needs:

Volume: Optimize packaging sizes to match your application scale.
Concentration: Customize concentrations for optimal performance.
Formulation: Available in liquid and lyophilized formats to suit your protocols.

Quality

Individual kit components are tested before and after formulation and post-lyophilization.

Functional assay
The performance of the reconstituted LyoBead is confirmed by an RPA reaction.

Frequently Asked Questions

What is Recombinase Polymerase Amplification (RPA) and how does it work?

Recombinase polymerase amplification, or RPA, is a remarkable isothermal DNA amplification method that does not need thermal cyclers, operates at 37-42 ºC, and provides exponential amplification of the targets.

The RPA process begins when recombinase proteins bind to primers and search for matching sequences in double-stranded DNA, enabling strand invasion and stabilization by single-stranded binding proteins. After the recombinase disassembles, a strand-displacing DNA polymerase elongates the primer, and repeated cycles of this reaction lead to exponential DNA amplification. For more information, refer to our comprehensive guide to the mechanisms and methods of RPA.

What types of samples and targets can I use with RPA Real-Time kits?

Our RPA Real-Time Kits are greatly suited for highly sensitive, real-time quantification of DNA targets under 30 minutes for point-of-care and field-based applications with strong signal clarity and minimal hands-on time. A wide range of sample types are compatible with the RPA Real-Time Kits, including crude extracts in addition to purified nucleic acids.

What materials and equipment do I need for RPA?

To run a real-time RPA reaction with our kits, you’ll need:

A pair of forward and reverse primers
An Exo probe for real-time signal detection with your choice of fluorophores (e.g., FAM, HEX, etc)
Nuclease-free water
Pipettes with sterile tips
Sterile tubes
Gloves

A heat block, water bath, or incubator is sufficient for a controlled, constant reaction temperature between 37 ºC and 42 ºC. A regular PCR thermal cycler works, too.

For real-time detection, you’ll need a fluorescence signal detection instrument. Our RPA Real-Time Kits are compatible with a wide range of such instruments, including regular real-time PCR systems. If you choose to use endpoint detection methods for the RPA Real-Time reaction products, common options include:

Colorimetric detection, commonly adopted in lateral flow assays
Electrophoresis-based detection
Other endpoint detection methods suited to your experimental design

Can I use PCR primers for RPA assays?

For optimal results, it is recommended to extend the length of target-specific PCR primers for the use in RPA experiments.

Optimal real-time RPA primers for our kits should be between 30 and 35 nt with a GC content between 30 and 60%. They should not be longer than 45 nt. Hence, RPA primers are slightly longer than common PCR primers (usually 18 to 30 nt). Considerations for reducing secondary structures (e.g., hairpins) and primer-dimer formations in primer design apply to RPA primers, too.

Please also design the real-time RPA primers so that the amplicons are between 110 and 200 bp, and should not exceed 400 bp, to achieve efficient and robust amplification.

Please design the Exo probe so that it has a length between 48 and 52 nt. Due to the use of such a probe for real-time measurement, our RPA Real-Time Kits have a slightly longer amplicon recommendation than the RPA Endpoint Kits.

What primer concentration should I use for RPA?

Optimal final primer and probe concentrations should be determined through titration testing for each specific assay. A good starting point is usually 0.4 – 0.5 µM of each primer and ~ 0.12 µM of the probe in a 50-µL reaction, and one should adjust based on the assay performance.

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