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A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.
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- A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.
- Nucleic acids research 2011, 39 (8):e55
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|dc.contributor.author||Robertson, Adam B||en|
|dc.contributor.author||Dahl, John A||en|
|dc.contributor.author||Vågbø, Cathrine B||en|
|dc.contributor.author||Krokan, Hans E||en|
|dc.identifier.citation||Nucleic acids research 2011, 39 (8):e55||en|
|dc.description.abstract||Recently, 5-hydroxymethylcytosine (5hmC) was identified in mammalian genomic DNA. The biological role of this modification remains unclear; however, identifying the genomic location of this modified base will assist in elucidating its function. We describe a method for the rapid and inexpensive identification of genomic regions containing 5hmC. This method involves the selective glucosylation of 5hmC residues by the β-glucosyltransferase from T4 bacteriophage creating β-glucosyl-5-hydroxymethylcytosine (β-glu-5hmC). The β-glu-5hmC modification provides a target that can be efficiently and selectively pulled down by J-binding protein 1 coupled to magnetic beads. DNA that is precipitated is suitable for analysis by quantitative PCR, microarray or sequencing. Furthermore, we demonstrate that the J-binding protein 1 pull down assay identifies 5hmC at the promoters of developmentally regulated genes in human embryonic stem cells. The method described here will allow for a greater understanding of the temporal and spatial effects that 5hmC may have on epigenetic regulation at the single gene level.||en|
|dc.subject||VDP::Medisinske Fag: 700::Basale medisinske, odontologiske og veterinÃ¦rmedisinske fag: 710::Medisinsk genetikk: 714||en|
|dc.subject.mesh||Embryonic Stem Cells||en|
|dc.subject.mesh||Gene Expression Regulation||en|
|dc.subject.mesh||Promoter Regions, Genetic||en|
|dc.title||A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.||en|
|dc.contributor.department||Centre for Molecular Biology and Neuroscience and Institute of Medical Microbiology, Oslo University Hospital, Rikshospitalet, Norway.||en|
|dc.identifier.journal||Nucleic acids research||en|
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