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Chapters
- 00:05Title
- 01:20Genomic DNA Fragmentation
- 02:02Glucosyl Transfer: B-GT Reaction
- 02:49Biotinylation Reaction: Click Chemistry
- 03:30Capture of 5-hmC-containing DNA
- 04:51Results: Selective Capture of 5-hmC from Genomic DNA
- 06:01Conclusion
Described is a two-step labeling process using β-glucosyltransferase (β-GT) to transfer an azide-glucose to 5-hmC, followed by click chemistry to transfer a biotin linker for easy and density-independent enrichment. This efficient and specific labeling method enables enrichment of 5-hmC with extremely low background and high-throughput epigenomic mapping via next-generation sequencing.
Tags
Selective Capture5-hydroxymethylcytosineGenomic DNA5-methylcytosineBiological FunctionsGene ExpressionGenome IntegrityParental ImprintingX-chromosome InactivationDevelopmentAgingCancerMammalian CellsEmbryonic Stem CellsNeuronal CellsTET Family Iron (II)/α-ketoglutarate-dependent DioxygenasesMES CellsNormal HematopoiesisMalignanciesZygote DevelopmentBisulfite SequencingLabeling And CaptureBacteriophage EnzymeGlucose MoietyGlucosyltransferase
