Related Experiment Video
Updated: Dec 25, 2025
12:34
DNA Methylation: Bisulphite Modification and Analysis
Published on: October 21, 2011
106.1K
Quantitative Analysis of DNA Methylation by Bisulfite Sequencing
Vasily V Ashapkin1, Lyudmila I Kutueva2, Boris F Vanyushin2
1Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia. ashapkin@genebee.msu.ru.
Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2020
Summary
Epigenetic clocks use DNA methylation patterns to predict biological age. This study presents a direct sequencing protocol for precise measurement of DNA methylation at specific CpG sites, enhancing age prediction accuracy.
Area of Science:
- Epigenetics and Molecular Biology
- Genomics and Bioinformatics
- Mammalian Aging Research
Background:
- Deoxyribonucleic acid (DNA) methylation patterns change with mammalian aging.
- Specific CpG site methylation levels correlate strongly with chronological age.
- These age-correlated CpG sites form the basis of epigenetic clocks for biological age prediction.
Purpose of the Study:
- To present a novel protocol for assessing individual CpG site DNA methylation levels.
- To enable precise quantitative measurement of DNA methylation in target sequences.
- To refine the accuracy and reliability of epigenetic clock applications.
Main Methods:
- Utilized bisulfite treatment of DNA to convert unmethylated cytosines.
- Employed polymerase chain reaction (PCR) to amplify target DNA sequences.
- Performed direct sequencing of PCR amplification products for methylation analysis.
Main Results:
- Successfully developed and validated a protocol for direct sequencing of bisulfite-converted DNA.
- Demonstrated the capability to quantitatively assess methylation levels at individual CpG sites.
- Established a reliable method for obtaining precise DNA methylation data for epigenetic clock construction.
Conclusions:
- The presented protocol offers a reliable and accurate method for measuring DNA methylation at specific CpG sites.
- This technique can enhance the precision of epigenetic clocks for biological age determination.
- Direct sequencing of bisulfite-converted DNA provides a robust approach for aging research and diagnostics.
