Cereal Disease Laboratory 1 article published in JoVE Genetics Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing Marisa E. Miller*1,2, Katie L. Liberatore*1,3, Shahryar F. Kianian1,3 1Cereal Disease Laboratory, United States Department of Agriculture-Agricultural Research Service, 2Department of Horticultural Science, University of Minnesota, 3Department of Plant Pathology, University of Minnesota The comparison and optimization of two plant organellar DNA enrichment methods are presented: traditional differential centrifugation and fractionation of the total gDNA based on methylation status. We assess the resulting DNA quantity and quality, demonstrate performance in short-read next-generation sequencing, and discuss the potential for use in long-read single-molecule sequencing.