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Methods Mol Biol. 2014;1176:23-31. doi: 10.1007/978-1-4939-0992-6_3.

Reduced representation bisulfite sequencing to identify global alteration of DNA methylation.

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Department of Pathology, Yale University School of Medicine, LH-214A, 310 Cedar Street, New Haven, CT, 06510, USA.


Reduced representation bisulfite sequencing is a cost-effective high-throughput sequencing-based method to obtain DNA methylation status at a single-nucleotide level. DNA methylation status is determined by utilizing DNA methylation-specific restriction enzymes to selectively amplify for genomic regions that are rich in methylated DNA. Although the method is genome-wide, DNA methyl sequencing does not require the sequencing of the whole genome, hence the name "reduced representation." However, a large majority of CpG islands are covered by reduced representation bisulfite sequencing allowing for the acquisition of comprehensive information of the methylation landscape in diseases like cancer. Data generated by this approach is typically reproducible and often covers between 65 and 75 % of the whole genome.

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