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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 18, 2026

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Published on: November 15, 2017

Simultaneous multiple target detection in real-time loop-mediated isothermal amplification.

Nathan A Tanner¹, Yinhua Zhang, Thomas C Evans

¹New England Biolabs, Ipswich, MA, USA.

|October 4, 2012

Summary

This summary is machine-generated.

Multiplex loop-mediated isothermal amplification (LAMP) enables simultaneous detection of multiple DNA targets in one reaction. This advancement enhances the utility of LAMP for rapid molecular diagnostics.

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Area of Science:

Molecular Biology
Biotechnology
Nucleic Acid Amplification

Background:

Loop-mediated isothermal amplification (LAMP) is a rapid, sequence-specific nucleic acid amplification method.
Current real-time LAMP detection is limited to single targets, restricting its diagnostic applications.

Purpose of the Study:

To develop a multiplex real-time detection method for LAMP capable of identifying multiple targets simultaneously.
To adapt LAMP primers for real-time fluorescence detection and assess the performance of new DNA polymerases.

Main Methods:

Modified standard LAMP primers with quencher-fluorophore duplexes for real-time fluorescent signal generation.
Tested the multiplex LAMP assay with 1-4 target sequences in a single reaction tube.
Evaluated the performance using Bst 2.0 and Bst 2.0 WarmStart DNA polymerases for faster amplification.

Main Results:

Achieved real-time detection of 1-4 targets in a single LAMP reaction.
Demonstrated high reproducibility and sensitivity, detecting below 100 copies of human genomic DNA.
Observed a 7-order of magnitude dynamic range and 50% faster amplification with new DNA polymerases.

Conclusions:

Developed a novel multiplex real-time LAMP assay for simultaneous detection of multiple nucleic acid targets.
The new method is sensitive, reproducible, and robust, significantly expanding LAMP's diagnostic potential.
Integration with advanced isothermal DNA polymerases enhances speed and broadens the utility of LAMP in molecular diagnostics.