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Selective Capture of 5-hydroxymethylcytosine from Genomic DNA
JoVE Revista
Biología
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JoVE Revista Biología
Selective Capture of 5-hydroxymethylcytosine from Genomic DNA
DOI:

06:26 min

October 05, 2012

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Capítulos

  • 00:05Título
  • 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

Summary

Traducción Automática

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.

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