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DNA Methylation: Bisulphite Modification and Analysis
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Biologie
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Journal JoVE Biologie
DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

DOI:
10.3791/3170-v

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12:34 min

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October 21, 2011

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

Chapitres

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

Summary

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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

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