In multiplex molecular testing, multiplex melting curve analysis is a key technology for overcoming the limitations of fluorescence channels and detecting multiple targets in a single tube. It is widely used in SNP genotyping, point-mutation identification, multiplex pathogen identification and probe performance verification. TaqMan hydrolysis probes and molecular beacon stem-loop probes use different fluorescence mechanisms, placing different demands on an instrument's optical system, temperature control precision and resistance to interference.

Rocgene's Archimed real-time PCR series combines patented scanning technology, precise temperature control and flexible multi-channel configurations to support multiplex melting curve experiments with molecular beacons, TaqMan probes and other probe systems. The series supports clinical diagnosis, pathogen screening and genotyping.

01 Two Major Probe Systems

Key Differences in Melting Curve Experiments

Multiplex melting curve analysis distinguishes multiple targets within a single fluorescence channel by using differences in the melting temperatures (Tm) of probes or targets. This enables efficient detection of more targets with fewer channels. Common challenges include fluorescence cross-talk that can cause false positives, interference from unwanted peaks, Tm shifts and distorted peak shapes caused by uneven temperatures, and weak signals from low-concentration samples.

TaqMan Probe Approach

Linear hydrolysis probes offer stable quantification and straightforward design. Challenges include broader melting peaks, lower resolution, susceptibility to background fluorescence, and limited specificity when distinguishing targets with similar Tm values. This approach is suited to routine multiplex quantification and rapid assay development.

Molecular Beacon Probe Approach

Reversible stem-loop probes offer very low background fluorescence, sharp peaks and strong single-base discrimination. They require highly precise temperature control, effective suppression of optical cross-talk and a high signal-to-noise ratio. Standard instruments may not meet the demands of such high-precision experiments. This approach is suited to precise, highly multiplexed testing.

Across the Archimed series, the hardware platform is designed to bring out the strengths of both probe systems and address common problems in melting curve experiments at their source.

02 Understanding the Complete Product Range

Melting Curve Performance and Application Scenarios

Mini Series | Portable Instruments for Rapid, Low-Throughput Testing

• Positioning: Compact, portable 16-well instruments with rapid heating and cooling, designed for both on-site screening and routine laboratory experiments.

• Key hardware specifications: Four fluorescence channels as standard. Mini 16 offers heating and cooling rates of up to 5.4–8°C/s and temperature control accuracy of ±0.2°C. Mini Pro reaches up to 11°C/s, with temperature control accuracy of ±0.15°C. Both instruments are small and lightweight, supporting mobile use.

• Melting curve and probe compatibility: The optical and temperature control systems support multiplex melting curve experiments with TaqMan and molecular beacon probes, producing regular peak shapes with few unwanted peaks. Targets with Tm differences of ≤1°C can be distinguished, meeting the needs of routine sample screening and basic probe verification.

• Applications: Low-throughput rapid clinical screening in primary healthcare facilities, community clinics, border quarantine and mobile emergency testing. Research applications include university teaching, exploratory experiments in small laboratories, rapid verification of field samples and small-scale preliminary studies.

X Series | Versatile 96-Well Instruments for Multiplex Melting Curve Analysis

• Positioning: High-throughput instruments with low cross-talk and a patented time-resolved optical system, designed for multiplex melting curve experiments.

• Key hardware specifications: An open-structure temperature control module with edge compensation provides temperature uniformity. Time-resolved scanning helps prevent cross-talk, with low background noise in molecular beacon experiments. Heating and cooling rates are 3.6–6°C/s, with consistent data across the plate.

• Melting curve and probe compatibility: Compatible with both TaqMan and molecular beacon probes. Standard melting peak shapes without shifts enable precise discrimination of targets with Tm differences of ≤1°C within a single channel. Strong single-base discrimination supports high-resolution melting (HRM), gene mutation analysis and development of complex multiplex assays. The six-channel X6 supports higher levels of multiplex detection in a single tube.

• Applications: Screening at centres for disease control and prevention, routine hospital testing, university research, pharmaceutical assay development, SNP genotyping, multiplex identification of complex pathogen panels and development of molecular beacon melting curve assays. Research uses include high-precision molecular beacon analysis, assay method development and mutation-site discrimination.

X Pro Series | Flagship 96-Well Instruments for Demanding Applications

• Positioning: A comprehensive upgrade of the X series optical and temperature control platform, with a focus on stability and expandability.

• Key hardware specifications: Upgraded matrix optical-fibre scanning, six fluorescence channels as standard, and expansion to eight channels. Temperature uniformity is ≤±0.1°C. Faster scanning, low signal-to-noise characteristics, high detection resolution and improved long-term operating stability support demanding workflows.

• Melting curve and probe compatibility: Supports both major probe systems and produces standard melting curves even with low-copy samples and complex templates, with highly reproducible peak shapes.

• Applications: PCR laboratories in tertiary hospitals, independent clinical laboratories, advanced tumour-targeted mutation genotyping, genetic disease variant testing, registration validation of IVD multiplex melting curve kits and high-standard multiplex screening for infectious diseases.

03 An Ideal Choice for Multiplex Melting Curve Experiments

From the high specificity of molecular beacons to the stable quantification of TaqMan probes, and from flexible research workflows to regulated clinical applications, Rocgene's Archimed series addresses the challenges of multiplex melting curve experiments through low cross-talk, high sensitivity, precise temperature control and broad probe compatibility.

Whether the goal is single-base resolution in basic research or efficient, precise clinical diagnosis, the Archimed series provides stable, reliable testing support for molecular biology research and precision medicine.