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Methylated DNA Immunoprecipitation

Methylated DNA Immunoprecipitation

Published on: January 2, 2009

Methylome analysis using MeDIP-seq with low DNA concentrations.

Oluwatosin Taiwo¹, Gareth A Wilson, Tiffany Morris

¹University College London (UCL) Cancer Institute, University College London, London, UK. t.taiwo@ucl.ac.uk

Nature Protocols

|March 10, 2012

Summary

This summary is machine-generated.

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This study presents an improved DNA methylation analysis protocol, Methylated DNA Immunoprecipitation-sequencing (MeDIP-seq), requiring significantly less DNA. This method enables accurate methylome generation from rare cells, advancing epigenetic research.

Area of Science:

Epigenetics
Genomics
Molecular Biology

Background:

DNA methylation is a key epigenetic regulator of biological processes.
Understanding its role in phenotypic plasticity requires genome-wide analysis.
Existing techniques often demand high DNA input, limiting studies on rare cell populations.

Purpose of the Study:

To develop an improved Methylated DNA Immunoprecipitation-sequencing (MeDIP-seq) protocol.
To reduce the required genomic DNA input for accurate methylome generation.
To enable MeDIP-seq analysis on limited samples, such as rare bone marrow cells.

Main Methods:

Optimization of the MeDIP-seq protocol for reduced DNA input.
Validation of specificity and enrichment across a range of DNA concentrations.

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Last Updated: May 24, 2026

Methylated DNA Immunoprecipitation

Methylated DNA Immunoprecipitation

Published on: January 2, 2009

Methyl-binding DNA capture Sequencing for Patient Tissues

Methyl-binding DNA capture Sequencing for Patient Tissues

Published on: October 31, 2016

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

Application of the protocol to generate methylomes from rare bone marrow cells.

Main Results:

The improved MeDIP-seq protocol uses 100-fold less genomic DNA than standard methods.
Comparable specificity (>97%) and enrichment (>100-fold) were achieved with 5-50 ng of DNA.
Successful methylome generation was demonstrated using only 160-300 ng of DNA from rare bone marrow cells.
The entire protocol, from DNA extraction to library generation, can be completed in 3-5 days.

Conclusions:

The optimized MeDIP-seq protocol significantly lowers DNA input requirements.
This advancement facilitates genome-wide DNA methylation analysis in samples with limited cellular material.
The protocol offers a cost-effective, accurate, and efficient method for methylome generation, supporting research into phenotypic plasticity and rare cell epigenetics.