Paramecium flow cytometry-based nucleus purification. Paramecium tetraurelia strain:51

We report a novel experimental approach for genome sequencing that uses flow cytometry to sort nuclei to high purity from complex biological samples. We employed this approach to sort subpopulations of nuclei in a mixture of nuclei with different sizes, or small and rare nuclei, isolated from the unicellular eukaryote Paramecium tetraurelia. We validated the method by flow imaging and high throughput DNA sequencing. This method enabled us to perform de novo genome assembly. This method to obtaining highly purified nuclei from complex samples should facilitate the analysis and sorting of minority subpopulations of nuclei, if a specific fluorophore, or specific characteristics of the nuclei, such as nucleus size or shape, or DNA content, can be associated with this subpopulation.