Abstract
Here we describe a method for genome wide investigation of methylation and hydroxymethylation status of cytosines. This protocol is an improvement of the HELP-tagging protocol previously described by Suzuki et al. It involves the glucosylation of 5-hydroxymethylcytosines (5-hmC) with β-glucosyl transferase (β-GT), thus rendering them resistant to digestion by MspI. Parallel digestion of β-GT treated samples with MspI, untreated sample with MspI and another untreated sample with HpaII, followed by adapter ligation, parallel sequencing and bioinformatics processing results in a differential display of MspI digestion sites that allows the determination of the distribution of 5-methylcytosines (5-mC) and 5-hmC at these sites.
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Battachariyya, S., Tytarenko, R., Heuck, C., Greally, J., Verma, A. (2018). Genome Wide Mapping of Methylated and Hydroxyl-Methylated Cytosines Using a Modified HpaII Tiny Fragment Enrichment by Ligation Mediated PCR Tagged Sequencing Protocol. In: Heuck, C., Weinhold, N. (eds) Multiple Myeloma. Methods in Molecular Biology, vol 1792. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7865-6_12
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DOI: https://doi.org/10.1007/978-1-4939-7865-6_12
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