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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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Related Experiment Video

Updated: May 23, 2026

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
07:27

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

Real time quantitative amplification detection on a microarray: towards high multiplex quantitative PCR.

Anke Pierik1, Marius Boamfa, Martijn van Zelst

  • 1Philips Research, High Tech Campus 11, 5656 AE Eindhoven, The Netherlands. anke.pierik@philips.com

Lab on a Chip
|April 5, 2012
PubMed
Summary
This summary is machine-generated.

A novel method enhances quantitative amplification detection beyond the typical 6-plex limit of quantitative real-time polymerase chain reaction (qrtPCR). This technique integrates solution amplification with real-time microarray hybridization for higher multiplexing capabilities.

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Last Updated: May 23, 2026

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Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
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Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Quantitative real-time polymerase chain reaction (qrtPCR) is a standard tool in research and diagnostics.
  • Current qrtPCR methods are limited to approximately 6-plex due to fluorophore spectral overlap.
  • Higher multiplexing is desired for increased efficiency and throughput in molecular assays.

Purpose of the Study:

  • To develop a novel method for quantitative amplification detection with enhanced multiplexing capabilities.
  • To overcome the limitations of spectral overlap in traditional qrtPCR.
  • To enable higher throughput molecular analysis in a single closed-chamber system.

Main Methods:

  • Integration of amplification in solution with real-time monitoring via hybridization to a microarray.
  • Utilized labeled primers and a compact, confocal scanner with miniaturized optics.
  • Combined scanning and heating using a flat thermocycler within a single closed chamber.

Main Results:

  • Demonstrated feasibility of the method in singleplex assays.
  • Achieved analytical sensitivity comparable to routine qrtPCR.
  • The system allows for quantitative amplification detection without PCR product manipulation.

Conclusions:

  • The presented method enables quantitative amplification detection at higher multiplexing levels than conventional qrtPCR.
  • This approach overcomes spectral overlap limitations by combining solution-based amplification with microarray detection.
  • The system offers a sensitive and efficient platform for advanced molecular diagnostics and research.