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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
Beskrives en totrins-mærkning ved anvendelse β-glucosyltransferase (β-GT) for at overføre et azid-glucose til 5-HMC, efterfulgt af klik kemi til at overføre en biotin linker til nem og densitet-uafhængig berigelse. Denne effektive og specifikke mærkning metode gør det muligt berigelse af 5-HMC med ekstremt lav baggrund og højkapacitetsidentifikation epigenomic kortlægning via næste generation sekventering.
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
