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Single Oocyte Bisulfite Mutagenesis
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JoVE Journal Biology
Single Oocyte Bisulfite Mutagenesis

Single Oocyte Bisulfite Mutagenesis

DOI:
10.3791/4046-v

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13:18 min

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June 27, 2012

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, ,
1Department of Obstretrics & Gynaecology,Schulich School of Medicine and Dentistry, University of Western Ontario, 2Department of Biochemistry,Schulich School of Medicine and Dentistry, University of Western Ontario, 3Children’s Health Research Institute

Chapters

  • 00:05Title
  • 01:22Oocyte Collection
  • 03:09Agarose Embedding and Lysis
  • 04:41Bisulfite Mutagenesis
  • 07:00First and Second Round PCR Amplification
  • 09:09TA Cloning and Colony PCR
  • 10:31Results: Electrophoretic Analysis of Single Oocyte Bisulfite Mutagenesis
  • 12:40Conclusion

Summary

Automatic Translation

Automatic Translation

Bisulfite mutagenesis is the gold standard for analyzing DNA methylation. Our modified protocol allows for DNA methylation analysis at the single-cell level and was specifically designed for individual oocytes. It can also be used for cleavage-stage embryos.

Tags

Single Oocyte Bisulfite MutagenesisEpigeneticsChromatin ModificationsGene AccessibilityGene TranscriptionDNA MethylationRepressive MarkCpG DinucleotidesMethyl GroupRepressive ProteinsHistone ModificationsDevelopmental ProgrammingCell DifferentiationRetroviral ElementsX-chromosome InactivationGenomic ImprintingBisulfite MutagenesisSodium BisulfiteDeaminationUracilsPCR AmplificationSequencingConversion EventsThyminesMethylated CytosinesNucleotide LevelAgarose Bead

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