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MethyLight and Digital MethyLight.

Mihaela Campan¹, Daniel J Weisenberger², Binh Trinh³

¹Clinical Sciences Building, University Hospital, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.

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

|December 11, 2017

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MethyLight is a quantitative PCR method for detecting DNA methylation. This technique, combined with quality controls and Digital PCR, enables sensitive detection of methylated DNA for disease screening.

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

DNA methylation is a critical epigenetic modification involved in gene regulation.
Detecting low-frequency methylated DNA regions is challenging due to high background levels of unmethylated DNA.

Purpose of the Study:

To detail the MethyLight method for sensitive and quantitative detection of DNA methylation.
To introduce and validate quality control reactions for MethyLight analysis.
To describe Digital MethyLight for ultra-sensitive detection of methylated DNA.

Main Methods:

MethyLight combines methylation-specific priming and fluorescent probing with real-time PCR.
Three types of quality control reactions (QC-1, QC-2, QC-3) were designed and tested.
Digital PCR was integrated with MethyLight to create Digital MethyLight.

Keywords:

Bisulfite Cancer DNA Methylation Digital PCR Epigenetics Methylation-specific PCR Quantitative Real-time PCR TaqMan

Related Experiment Videos

Main Results:

MethyLight accurately quantifies low-frequency methylated DNA regions.
Quality control reactions effectively monitor sample integrity, bisulfite conversion efficiency, and treatment completeness.
Digital MethyLight demonstrated highly sensitive detection of individual methylated DNA molecules.

Conclusions:

MethyLight provides a robust and quantitative approach for DNA methylation analysis.
Integrated quality controls enhance the reliability of MethyLight results.
Digital MethyLight offers a promising tool for sensitive disease detection and screening.