Journal
/
Biology
/
DNA Methylation: Bisulphite Modification and Analysis
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

DOI:
10.3791/3170-v

•

12:34 min

•

October 21, 2011

•

, , ,
1Epigenetics Group, Cancer Research Program,Garvan Institute of Medical Research, 2St Vincent’s Clinical School,University of NSW

Chapters

  • 00:05Title
  • 01:52DNA Preparation and Denaturation
  • 02:57Bisulphite Deamination
  • 06:38PCR Amplification
  • 09:08Results of DNA Methylation Analysis
  • 12:15Conclusion

Summary

Automatic Translation

Automatic Translation

The gold standard for DNA methylation analysis is genomic sequencing of bisulphite converted DNA. This method takes advantage of the increased sensitivity of cytosine compared with 5-methylcytosine (5-MeC) to bisulphite deamination under acidic conditions. Unmethylated cytosines can be distinguished from methylated cytosines after PCR amplification of the target genomic DNA.

Tags

DNA MethylationBisulphite ModificationAnalysisEpigeneticsGene FunctionDNA SequenceDNA Methylation Patterns5-MeCGene Expression MachineryCpG DinucleotidesGenomic SequencesGene PromotersDNA Methylation AnalysisBisulphite ConversionCytosine DeaminationSodium Bisulphite

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/3170?article_type=v

PDMS Device Fabrication and Surface Modification
check_url/3170?article_type=v

A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity
check_url/3170?article_type=v

Determination of DNA Methylation of Imprinted Genes in Arabidopsis Endosperm
check_url/3170?article_type=v

Genetic Modification and Recombination of Salivary Gland Organ Cultures
check_url/3170?article_type=v

An Efficient Method for Quantitative, Single-cell Analysis of Chromatin Modification and Nuclear Architecture in Whole-mount Ovules in Arabidopsis
check_url/3170?article_type=v

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
check_url/3170?article_type=v

Reduced-gravity Environment Hardware Demonstrations of a Prototype Miniaturized Flow Cytometer and Companion Microfluidic Mixing Technology
check_url/3170?article_type=v

In vitro Methylation Assay to Study Protein Arginine Methylation
check_url/3170?article_type=v

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
check_url/3170?article_type=v

Correlating Gene-specific DNA Methylation Changes with Expression and Transcriptional Activity of Astrocytic KCNJ10 (Kir4.1)

Read Article