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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: Nov 3, 2025

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

Published on: October 7, 2025

Spatially localized amplification reaction with accelerated target conversion for sensitive microRNA detection.

Min Qing¹, Sheng Liang Chen¹, Jiao Zhou¹

¹School of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, China.

|June 2, 2021

Summary

We developed a spatially localized amplification reaction (SLAR) for sensitive microRNA-21 (miRNA-21) detection. This novel nanoprobe assay enhances signal gain and specificity, showing promise for early disease diagnosis.

Keywords:

Entropy-driven reaction Fluorescence assay MicroRNA Spatial-confinement effects Spatially localized amplification reaction

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Published on: August 3, 2011

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Last Updated: Nov 3, 2025

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

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Published on: October 7, 2025

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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

Area of Science:

Biochemistry
Nanotechnology
Molecular Biology

Background:

MicroRNA-21 (miRNA-21) is a biomarker for various diseases.
Sensitive and specific detection methods are crucial for early diagnosis.
Existing detection methods can be limited by reaction kinetics and specificity.

Purpose of the Study:

To develop a novel nanoprobe assay for sensitive and specific detection of miRNA-21.
To enhance reaction efficiency and signal gain using a spatially localized amplification reaction (SLAR).
To demonstrate the assay's potential for biomarker detection in cell lysates.

Main Methods:

Constructed a spatially localized amplification reaction (SLAR) nanoprobe by colocalizing entropy-driven reactions (EDR) on a DNA scaffold.
Utilized DNA self-assembly for probe fabrication.
Triggered interval EDR along the DNA scaffold by target miRNA-21, leading to fluorescence recovery.

Main Results:

The SLAR assay achieved a low detection limit of 6 pM for miRNA-21.
Demonstrated excellent specificity in distinguishing miRNA-21 from similar microRNAs.
Successfully detected miRNA-21 in MCF-7 and HeLa cell lysates.

Conclusions:

The SLAR nanoprobe offers a highly sensitive and specific method for miRNA detection.
Spatial confinement of reactions on a nanoscale scaffold accelerates target conversion and signal generation.
The assay holds significant promise for the sensitive detection of biomarkers in early disease diagnosis.