Distinguishing DNA Modifications

Chromatin states play important roles in cellular differentiation and stem cell dynamics. Specifically, cytosine modifications are known to influence histone remodeling and fine tune gene expression. Changes in these epigenetic modifications have also been linked to neurodegenerative diseases and cancer. Increasing evidence suggests that different cytosine modifications such as 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) are functionally distinct, and should be studied as such. However, traditional bisulfite sequencing cannot distinguish between these modifications, and yields a combined result of unknown proportions. To address this problem, Cambridge Epigenetix developed TrueMethyl™ oxidative bisulfite (oxBS) technology, which enables accurate quantification of 5mC and 5hmC in parallel. TrueMethyl™ is based upon the oxBS-Seq methodology developed by Prof. Shankar Balasubramanian, who previously co-invented the Solexa / Illumina sequencing platform. TrueMethyl™ oxBS technology enables rapid and robust quantitative measurements of 5mC and 5hmC in parallel at single-base resolution.

 

Achieve complete measurements of methylation with oxBS. The schematic (Left) shows classic bisulfite conversion which creates a library that detects both 5mC and 5hmC together. Processing with the oxidation of 5hmC (Right) generates a bisulfite- convertible base that leads to detection of only 5mC. Differences between the libraries can then be used to deduce the sites of 5hmC modifications.

 


Return to TrueMethyl Homepage.
Learn more about 5-methylcytosine here.
Learn more about 5-hydroxymethylcytosine here.