Reduced representation bisulfite sequencing (RRBS) is an effective technique for profiling genome-wide DNA methylation patterns in eukaryotes. RRBS couples size selection, bisulfite conversion, and second generation sequencing to enrich for CpG-dense regions of the genome. The progressive improvement of second-generation sequencing technologies and reduction in cost provided an opportunity to examine the DNA methylation patterns of multiple genomes. Here, we describe a protocol for sequencing multiple RRBS libraries in a single sequencing reaction to generate base-resolution methylomes. Furthermore, we provide a brief guideline for base-calling and data analysis of multiplexed RRBS libraries. These strategies will be useful to perform large-scale, genome-wide DNA methylation analysis.
CITATION STYLE
Chatterjee, A., Rodger, E. J., Stockwell, P. A., Le Mée, G., & Morison, I. M. (2017). Generating multiple base-resolution DNA methylomes using reduced representation bisulfite sequencing. In Methods in Molecular Biology (Vol. 1537, pp. 279–298). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6685-1_16
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