Abstract：
:To uncover the function of and interplay between the mammalian cytosine modifications 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), new techniques and advances in current technology are needed. To this end, we have developed oxidative bisulfite sequencing (oxBS-seq), which can quantitatively locate 5mC and 5hmC marks at single-base resolution in genomic DNA. In bisulfite sequencing (BS-seq), both 5mC and 5hmC are read as cytosines and thus cannot be discriminated; however, in oxBS-seq, specific oxidation of 5hmC to 5-formylcytosine (5fC) and conversion of the newly formed 5fC to uracil (under bisulfite conditions) means that 5hmC can be discriminated from 5mC. A positive readout of actual 5mC is gained from a single oxBS-seq run, and 5hmC levels are inferred by comparison with a BS-seq run. Here we describe an optimized second-generation protocol that can be completed in 2 d.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Booth MJ,Ost TW,Beraldi D,Bell NM,Branco MR,Reik W,Balasubramanian Sdoi
10.1038/nprot.2013.115subject
Has Abstractpub_date
2013-10-01 00:00:00pages
1841-51issue
10eissn
1754-2189issn
1750-2799pii
nprot.2013.115journal_volume
8pub_type
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