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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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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
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Quantifying mRNA levels across tissue sections with 2D-RT-qPCR.

Michael Armani1, Michael A Tangrea, Benjamin Shapiro

  • 1Fischell Department of Bio-Engineering, University of Maryland, College Park, MD 20742, USA.

Analytical and Bioanalytical Chemistry
|May 12, 2011
PubMed
Summary

This study introduces two-dimensional RT-qPCR (2D-RT-qPCR), a novel method for quantifying mRNA in tissue sections. This integrated platform simplifies spatial RNA analysis and gene expression studies in disease research.

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Last Updated: Jun 2, 2026

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10:28

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MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)
08:30

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)

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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

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Measuring mRNA levels is crucial for understanding gene function and disease mechanisms.
  • Current methods for quantifying low-abundance mRNA often involve multiple platforms, complicating spatial information preservation.
  • Spatial transcriptomics requires preserving RNA location during analysis.

Purpose of the Study:

  • To introduce a novel, integrated method for quantifying RNA in tissue sections.
  • To simplify the workflow for spatial mRNA analysis.
  • To demonstrate the feasibility of a new high-throughput tissue analysis platform.

Main Methods:

  • Developed two-dimensional RT-qPCR (2D-RT-qPCR) on a single integrated platform.
  • Utilized a multi-well plate format for macrodissection of tissue sections, preserving spatial RNA location.
  • Implemented an in-plate lysis and nucleic acid purification protocol followed by RT-qPCR.

Main Results:

  • Demonstrated the feasibility of 2D-RT-qPCR across various tissue types.
  • Successfully quantified RNA in tissue sections while preserving spatial information.
  • Eliminated the need for physical tissue homogenization.

Conclusions:

  • 2D-RT-qPCR offers a streamlined, single-platform approach for spatial RNA quantification.
  • The method simplifies complex workflows, enabling efficient gene expression analysis.
  • This technology has potential as a high-throughput platform for tissue analysis.