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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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Updated: Mar 26, 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

Microfluidics-Based PCR for Fusion Transcript Detection.

¹Department of Pathology, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA. hchen7@mdanderson.org.

Methods in Molecular Biology (Clifton, N.J.)

|February 5, 2016

Summary

This summary is machine-generated.

Microfluidic polymerase chain reaction (PCR) enables simultaneous testing of multiple samples and assays in nanoliter volumes. This technology is crucial for analyzing limited DNA/RNA samples, particularly for leukemia fusion transcript screening.

Keywords:

Dynamic array Integrated fluidic circuits Microfluidics-based PCR Real-time PCR

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Area of Science:

Biotechnology
Molecular Biology
Medical Diagnostics

Background:

Microfluidic technology enables the creation of intricate networks of submillimeter fluidic channels and reservoirs.
Microfluidic-based polymerase chain reaction (PCR) facilitates automated, simultaneous multiplexing of samples and assays.
Nanoliter reaction volumes are achievable, ideal for precious samples with limited DNA and RNA.

Purpose of the Study:

To describe the application of microfluidics-based PCR for the simultaneous screening of leukemia fusion transcripts.
To highlight the advantages of microfluidic PCR in handling limited sample volumes and multiplexed assays.

Main Methods:

Utilizing microfluidic devices for polymerase chain reaction (PCR).
Implementing automated valve systems for controlled fluidic channel and chamber coordination.
Performing multiplexed assays for the detection of 14 known fusion transcripts.

Main Results:

Demonstrated the capability of microfluidic PCR for simultaneous screening of multiple targets.
Showcased the efficiency of nanoliter-volume reactions for genetic analysis.
Successfully applied the technology for leukemia-specific fusion transcript identification.

Conclusions:

Microfluidics-based PCR is a powerful tool for efficient and simultaneous genetic screening.
This technology significantly benefits leukemia diagnostics by enabling rapid analysis of fusion transcripts from limited samples.