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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: Jun 3, 2026

Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

Published on: June 11, 2016

Epiallele quantification using molecular inversion probes.

Ramkumar Palanisamy¹, Ashley R Connolly, Matt Trau

¹Centre for Biomarker Research and Development, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane Qld, 4072 Australia.

Analytical Chemistry

|March 16, 2011

Summary

This summary is machine-generated.

DNA methylation patterns are key cancer biomarkers. A new assay using molecular inversion probes and flow cytometry rapidly and accurately quantifies epialleles in genomic DNA samples.

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Area of Science:

Genomics
Epigenetics
Biomarker Discovery

Background:

DNA methylation is a critical epigenetic modification.
Aberrant DNA methylation patterns are hallmarks of various cancers.
Analyzing epialleles in heterogeneous samples is challenging.

Purpose of the Study:

To develop a novel assay for comprehensive epiallele analysis.
To quantify DNA methylation at specific CpG sites with high resolution.
To enable rapid and sensitive detection of cancer biomarkers.

Main Methods:

Utilized molecular inversion probes for targeted DNA enrichment.
Applied bisulphite treatment to distinguish methylated and unmethylated cytosines.
Employed multiplexable microbead DNA biosensors with flow cytometry for readout.

Main Results:

Successfully exposed and quantified epialleles in bisulphite-treated genomic DNA.
Achieved rapid quantification of different CpG dinucleotides.
Demonstrated high resolution, sensitivity, specificity, and a large dynamic range.

Conclusions:

The developed assay provides a powerful tool for epiallele analysis.
This method facilitates accurate biomarker discovery for cancer diagnosis.
The assay offers a sensitive and efficient approach for epigenetic profiling.