Epigenetic Approaches in Non-Model Plants
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Reduced representation bisulfite sequencing is an emerging methodology for evolutionary and ecological genomics and epigenomics research because it provides a cost-effective, high-resolution tool for exploration and comparative analysis of DNA methylation and genetic variation. Here we describe how digestion of genomic plant DNA with restriction enzymes, subsequent bisulfite conversion of unmethylated cytosines, and final DNA sequencing allow for the examination of genome-wide genetic and epigenetic variation in plants without the need for a reference genome. We explain how the use of several combinations of barcoded adapters for the creation of highly multiplexed libraries allows the inclusion of up to 144 different samples/individuals in only one sequencing lane.
Key wordsEpigenetics DNA methylation Bisulfite treatment epiGBS Next-generation sequencing Restriction enzymes Multiplexing Reduced representation sequencing
This work was supported by funding from the National Science Foundation (U.S.A.) IOS-1556820 (to C.L.R.), and through the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska Curie grant agreement No. 704141 (to M.T.B.).
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