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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...

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Related Experiment Video

Updated: Jun 16, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
08:23

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

High-throughput droplet PCR.

Amelia L Markey1, Stephan Mohr, Philip J R Day

  • 1School of Chemical Engineering and Analytical Sciences, Manchester Interdisciplinary Biocentre, University of Manchester, UK.

Methods (San Diego, Calif.)
|February 2, 2010
PubMed
Summary
This summary is machine-generated.

Miniaturized fluidic devices enable droplet-based polymerase chain reaction (PCR) for analyzing complex tissues. This high-throughput method enhances nucleic acid analysis by preventing contamination and sample loss, enabling single-cell studies.

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Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics
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Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics

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Droplet Digital TRAP (ddTRAP): Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction
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Droplet Digital TRAP (ddTRAP): Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction

Published on: May 3, 2019

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Last Updated: Jun 16, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
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06:38

Droplet Digital TRAP (ddTRAP): Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction

Published on: May 3, 2019

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Microfluidics

Background:

  • The polymerase chain reaction (PCR) revolutionized nucleic acid analysis.
  • Analyzing complex, heterogeneous tissues requires new methods for high-throughput, quantitative data.
  • Current methods face challenges in handling small sample volumes and preventing contamination.

Purpose of the Study:

  • To develop a microfluidic system for generating droplets for droplet-based PCR.
  • To enable massively parallelized, quantitative nucleic acid analysis from cellular components of tissues.
  • To facilitate high-throughput, single-cell analysis of nucleic acids.

Main Methods:

  • Utilized miniaturized fluidic devices for producing aqueous droplets in a two-phase flow.
  • Designed and produced a prototype microfluidic device for droplet generation and stable flow.
  • Investigated droplet characteristics for suitability in continuous flow microfluidic PCR.

Main Results:

  • Demonstrated routine and reliable production of droplets using the microfluidic device.
  • Showcased stable droplet flow, essential for continuous flow microfluidic PCR applications.
  • Highlighted the potential of droplets for reactant containment, minimizing contamination and sample loss.

Conclusions:

  • Droplet-based microfluidic systems offer a promising platform for advanced nucleic acid analysis.
  • This technology facilitates high-throughput, single-cell analysis from complex biological samples.
  • Optimized fabrication and calibration of microfluidic devices are crucial for robust performance.