5-methylcytosine (5-mC) is the predominant epigenetic mark in mammalian genomic DNA. 5-hydroxymethylcytosine (5-hmC) is a newly discoveredepigenetic modification that is presumably generated by oxidation of 5-mC by the TET family of cytosine oxygenases.1,2
Techniques exist that can identify 5-mC in genomic DNA, but the most commonly used method, bisulfite sequencing, is laborious and cannot distinguish between 5-mC from 5-hmC.3The kit distinguishes 5-mC from 5-hmC by the addition of glucose to the hydroxyl group of 5-hmC via an enzymatic reaction utilizing T4 β-glucosyltransferase (T4-BGT). When 5-hmC occurs in the context of CCGG, this modification converts a cleavable MspI site to a noncleavable one.An overview of the detection procedure is summarized in Figure 1.Control DNA Sequence5´-CAGTGAAGTTGGCAGACTGAGCCAGGTCCCACAGATGCAGTGACCGGAGT CATTGCCAAACTCTGCAGGAGAGCAAGGGCTGTCTATAGGTGGCAAGTCA-3´Control DNA substrates are synthetic 100 bp double stranded fragments containing a single MspI/HpaII site (CCGG). The three fragments are identical except for modification of the internal C in this site.FW Primer Sequence5´- CA GTG AAG TTG GCA GAC TGA GC -3´REV Primer Sequence5´- CTG ACT TGC CAC CTA TAG ACA GC -3´
This product is related to the following categories:
Methylome Analysis,
Epigenetic Analysis,
Next Generation Sequencing Library Preparation
This product can be used in the following applications:
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