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Optimized Analysis of DNA Methylation and Gene Expression from Small, Anatomically-defined Areas of the Brain
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Chapters
- 00:05Title
- 01:58Programming by Early Life Adversity
- 02:45Collecting Brain Tissue Punches
- 04:02Nucleic Acid Extraction from Brain Punches
- 06:08Bisulfite PCR
- 07:41Bisulfite Sequencing
- 10:57The Effect of ELS on the Expression and Methylation of AVP
- 12:35Conclusion
A streamlined workflow to study DNA methylation and gene expression changes upon early-life stress is shown. Starting from maternal separation of newborn mice and isolation of discrete brain tissues, we represent a protocol to simultaneously isolate DNA and RNA from brain tissue punches for subsequent bisulfite sequencing and RT-PCR analysis.
Tags
DNA MethylationGene ExpressionBrain AnalysisSmall Anatomically-defined AreasDietDrugsEarly Life AdversityLasting ChangesPhysiological PhenotypesBehavioural PhenotypesMetabolic DiseasesCardiovascular DiseasesMental DiseasesEnvironmental ProgrammingGene-environment DialogueHistone ModificationsMammalsCpG MethylationCellular HeterogeneityGene Expression And Methylation CorrelationEpigenetic ProgrammingMaternal Separation ParadigmMicropunches Isolation
