Document Detail


The GLIB technique for genome-wide mapping of 5-hydroxymethylcytosine.
MedLine Citation:
PMID:  23018194     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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:
William A Pastor; Yun Huang; Hope R Henderson; Suneet Agarwal; Anjana Rao
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-09-27
Journal Detail:
Title:  Nature protocols     Volume:  7     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-09-28     Completed Date:  2013-01-28     Revised Date:  2013-10-17    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1909-17     Citation Subset:  IM    
Affiliation:
La Jolla Institute for Allergy and Immunology, La Jolla, California, USA.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Biotinylation / methods
Cytosine / analogs & derivatives*,  analysis,  chemistry
Genomics / methods*
Glucose / chemistry
Molecular Sequence Data
Oxidation-Reduction
Periodic Acid / chemistry
Grant Support
ID/Acronym/Agency:
1 UL1 RR 025758-02/RR/NCRR NIH HHS; AI44432/AI/NIAID NIH HHS; HD065812/HD/NICHD NIH HHS; HL089150/HL/NHLBI NIH HHS; R01 AI044432/AI/NIAID NIH HHS; R01 CA151535/CA/NCI NIH HHS; R01 HD065812/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
10450-60-9/Periodic Acid; 1123-95-1/5-hydroxymethylcytosine; 50-99-7/Glucose; 71-30-7/Cytosine; 7790-28-5/metaperiodate
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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