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Real Time RT-PCR

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: May 30, 2026

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

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

Published on: August 3, 2011

High-throughput microfluidic single-cell RT-qPCR.

Adam K White¹, Michael VanInsberghe, Oleh I Petriv

¹Centre for High-Throughput Biology, University of British Columbia, Vancouver, BC, Canada V6T 1Z4.

Proceedings of the National Academy of Sciences of the United States of America

|August 3, 2011

Summary

This summary is machine-generated.

Researchers developed an integrated microfluidic device for scalable single-cell transcription analysis. This technology enables high-precision gene expression measurements with increased sensitivity and specificity for various biological applications.

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Area of Science:

Biotechnology
Molecular Biology
Microfluidics

Background:

Scalable analysis of transcription in single cells is a significant challenge in microfluidics.
Existing methods often lack the throughput and precision required for comprehensive single-cell studies.

Purpose of the Study:

To develop a fully integrated microfluidic device for high-precision, high-throughput single-cell gene expression analysis.
To demonstrate the device's capability in measuring microRNA (miRNA) expression, transcript coregulation, and single nucleotide variants.

Main Methods:

Development of an integrated microfluidic device performing cell capture, lysis, reverse transcription, and quantitative PCR (RT-qPCR).
Nanoliter volume processing for enhanced measurement precision and sensitivity.
Application to over 3,300 single-cell measurements.

Main Results:

The device successfully performed high-precision RT-qPCR on hundreds of single cells per run.
Nanoliter volume processing reduced noise, increased sensitivity, and provided single nucleotide specificity.
Demonstrated utility in analyzing miRNA expression, miRNA-target coregulation during stem cell differentiation, and breast cancer cell single nucleotide variants.

Conclusions:

The integrated microfluidic device represents a milestone in scalable single-cell transcription analysis.
The technology provides a robust platform for diverse on-chip single-cell transcription studies.
This advancement facilitates deeper understanding of cellular heterogeneity and gene regulation.