Duplex Assay
Duplex Assay - Related Videos
12:14
A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
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Cited by 14 •
2016
This manuscript describes a duplex digital PCR assay that can be used to simultaneously quantify Enterococcus spp. and the HF183 genetic markers as indicators of general and human-associated fecal contamination in recreational waters.
08:45
Duplex Digital PCR for Simultaneous Quantification of Dual Genetic Markers
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2025
In this video, we demonstrate the duplex digital PCR (ddPCR) technique — a modification of the traditional PCR technique that is useful in detecting two different genetic markers simultaneously. A single PCR reaction is partitioned into nanoliter-sized emulsified droplets that are independently amplified, and the detection of differently colored fluorescence amplification signals from the fraction of droplets is used to compute the initial concentration of the target sequences.
11:42
Iterative Optimization of DNA Duplexes for Crystallization of SeqA-DNA Complexes
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2012
Crystal structure of protein–DNA complexes can provide insight into protein function, mechanism, as well as, the nature of the specific interaction. Here, we report how to optimize the length, sequence and ends of duplex DNA for co-crystallization with Escherichia coli SeqA, a negative regulator of replication initiation.
04:10
Helicase Activity Measurement of a Target Protein Using Biotin-Labeled RNA Duplexes
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2025
In this video, we demonstrate the procedure to determine the helicase activity of a target protein to unwind the biotin-labeled dsRNA substrate. The activity of the enzyme was identified by analyzing the electrophoretic mobility shift, followed by a chemiluminescence assay using chemiluminescent enzyme-conjugated streptavidin.
10:41
Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound
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2021
The aim of this manuscript is to present a sonography-based method that allows in vivo imaging of blood flow in cerebral arteries in mice. We demonstrate its application to determine changes in blood flow velocities associated with vasospasm in murine models of subarachnoid hemorrhage (SAH).
FAQs
Distinguishable fluorescent labels or signal channels assign each biological target its own measurement pathway. During the shared reaction, the signal associated with one primer-probe set, antibody, or other recognition reagent can be interpreted separately from the second signal. This separation is central to duplex assay design because simultaneous detection is useful only when both target-specific readouts remain individually readable.
Running two targets together can conserve sample and reduce handling compared with performing separate reactions for each target. The combined format also supports higher throughput while retaining target-specific readouts. These advantages are especially relevant when sample quantity is limited or when many specimens must be processed for pathogen testing, genetic analysis, expression studies, or biomarker measurement.
An internal control gives the reaction an additional reference signal that is measured alongside the target of interest. In a duplex design, one channel can represent the target while the other represents the control, allowing both signals to be assessed under shared reaction conditions. This arrangement is useful when the assay must examine target detection together with a control measurement.
The recognition reagents must correspond to their intended biological targets, while the associated fluorescent labels or signal channels must remain distinguishable. Duplex assay formats may use two primer and probe sets, two antibody-based reagents, or other target-recognition combinations. Matching each reagent to a separate readable signal preserves independent interpretation within the same reaction.
A basic design begins by selecting two biological targets and assigning each one a compatible recognition system, such as primers and probes or antibodies. The two systems are then paired with distinguishable labels or signal channels and evaluated under shared reaction conditions. After the reaction, each signal is read separately so the two target-specific measurements remain interpretable.
This approach is useful when a study needs information about two biological signals from one sample. Applications described for duplex assays include identifying pathogens, analyzing genetic variants, measuring gene expression, and testing biomarkers. A design may also combine a target with an internal control. In each case, the format links simultaneous measurement with reduced sample use and handling.