Gel-free multiplexed reduced representation bisulfite sequencing for large-scale DNA methylation profiling |
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| Authors: | Patrick Boyle Kendell Clement Hongcang Gu Zachary D Smith Michael Ziller Jennifer L Fostel Laurie Holmes Jim Meldrim Fontina Kelley Andreas Gnirke Alexander Meissner |
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| Institution: | 1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA;2Harvard Stem Cell Institute, Cambridge, MA 02138, USA;3Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA;4Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA 02139, USA |
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| Abstract: | Sequencing-based approaches have led to new insights about DNA methylation. While many different techniques for genome-scale mapping of DNA methylation have been employed, throughput has been a key limitation for most. To further facilitate the mapping of DNA methylation, we describe a protocol for gel-free multiplexed reduced representation bisulfite sequencing (mRRBS) that reduces the workload dramatically and enables processing of 96 or more samples per week. mRRBS achieves similar CpG coverage to the original RRBS protocol, while the higher throughput and lower cost make it better suited for large-scale DNA methylation mapping studies, including cohorts of cancer samples. |
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