A subscription to JoVE is required to view this content. Sign in or start your free trial.
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
Hydrogensulfit mutagenese er den gyldne standard for analyse af DNA methylering. Vores modificeret protokol giver mulighed for at DNA-methylering analyse på enkelt-celle niveau, og er specielt designet til individuelle oocytter. Det kan også anvendes til spaltning-embryoer.
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
