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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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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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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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Multiplex Real-Time PCR Using Encoded Microparticles for MicroRNA Profiling.

Seungwon Jung1, Sang Kyung Kim2,3

  • 1Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology, Seoul, South Korea.

Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2017
PubMed
Summary
This summary is machine-generated.

This study introduces a novel multiplex quantitative real-time PCR (qPCR) method for analyzing multiple microRNA (miRNA) expression. The technique uses unique microparticles for high-fidelity profiling of numerous genetic targets in a single reaction.

Keywords:
Encoded particleHydrogelMicroRNAMultiplexReal-time PCR

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Multiplex quantitative real-time PCR (qPCR) is crucial for analyzing multiple genetic targets simultaneously.
  • The demand for high-throughput genetic analysis is rapidly increasing.
  • Existing methods face challenges in efficiently analyzing numerous targets in a single sample.

Purpose of the Study:

  • To develop an extensible qPCR method for multiplex microRNA (miRNA) expression analysis.
  • To enable high-fidelity signal analysis in multiplex real-time PCR.
  • To facilitate the quantitative profiling of tens of miRNAs in a single reaction.

Main Methods:

  • Utilized microparticles with immobilized primers as discrete reactors for qPCR.
  • Employed two-dimensional codes engraved into microparticles for individual particle identification.
  • Achieved high-fidelity signal analysis during multiplex real-time PCR.

Main Results:

  • Demonstrated efficient amplification of amplicons within microparticles, exceeding 95% efficiency.
  • Successfully profiled tens of miRNAs quantitatively in a single PCR reaction.
  • The method proved readily extensible for analyzing multiple miRNA targets.

Conclusions:

  • The developed microparticle-based qPCR offers a robust and extensible solution for multiplex miRNA expression analysis.
  • This technique allows for high-fidelity and efficient profiling of numerous genetic targets.
  • It represents a significant advancement in high-throughput genetic analysis.