The GLIB technique for genome-wide mapping of 5-hydroxymethylcytosine. Academic Article

abstract

• 5-Hydroxymethylcytosine (5hmC) is a newly discovered DNA base present at detectable levels in most mammalian cell types and tissues. It is generated by Tet-enzyme-mediated oxidation of 5-methylcytosine (5mC). 5hmC is important both because of its potential role in regulating gene expression and because it may be an intermediate in DNA demethylation. Here we describe a technique termed GLIB (glucosylation, periodate oxidation and biotinylation), which combines several enzymatic and chemical modification steps to attach biotin to 5hmC. Biotin-containing genomic DNA fragments are then enriched using streptavidin beads, eluted and sequenced. GLIB is capable of quantitatively tagging and precipitating fragments containing a single 5hmC molecule. Sample preparation and GLIB can be conducted in 2-3 d.

authors

• Huang, Yun

published proceedings

• Nat Protoc

author list (cited authors)

• Pastor, W. A., Huang, Y., Henderson, H. R., Agarwal, S., & Rao, A.

citation count

• 51

complete list of authors

• Pastor, William A||Huang, Yun||Henderson, Hope R||Agarwal, Suneet||Rao, Anjana

publication date

• January 2012

publisher

• Springer Nature  Publisher

published in

• Nature Protocols  Journal

keywords

• 5-Methylcytosine
• Base Sequence
• Biotinylation
• Cytosine
• Genomics
• Glucose
• Molecular Sequence Data
• Oxidation-Reduction
• Periodic Acid

Digital Object Identifier (DOI)

• 10.1038/nprot.2012.104

start page

• 1909

end page

• 1917

volume

• 7

issue

• 10

URL

• http://dx.doi.org/10.1038/nprot.2012.104