Fig. 5.

Flowchart for eccDNA research. The schematic comprehensively details eccDNA research, and some of the steps are not essential. In their research, researchers can select some of the steps based on the strengths and limitations of the methods listed in Table 1. Karyotyping and EMs, which are classic techniques in most labs, can qualitatively identify eccDNAs. Deep sequencing shows numerous strengths but is expensive. The process of eccDNA structure validation, which can be achieved by an integrated approach combining FISH and inverse PCR, is critical for studying eccDNA. Functional and mechanistic assays include transcriptomic, proteomic, and other approaches