In eccDNA sequencing, the overwhelming amounts of linear chromosomal DNA and circular mitochondrial DNA (mtDNA) in the DNA sample must be removed as they can significantly reduce the eccDNA reads. Arraystar eccDNA-seq linearizes the circular mtDNAs by restriction endonuclease cleavage and then digests the linearized mtDNA and linear chromosomal DNA by a linear DNA-specific exonuclease, leaving eccDNAs intact and enriched in the sample. The eccDNAs are amplified by Rolling Circle Amplification (RCA), greatly improving the eccDNA signals and data quality.
• Broad coverage: Simultaneous profiling of eccDNA(<10 kb) and ecDNA(>=10 kb).
• Maximal eccDNA enrichment: Removal of chromosomal and mitochondrial DNA and Rolling Circle Amplification of eccDNA to dramatically increase eccDNA reads.
• High reliability: Well established and optimized procedures to produce best possible results.
• Easy-to-use analyses: Provided with rich annotation, genome browser track visualization, publication-ready graphics for biologists and clinicians.
Watch Video> The Extraordinary eccDNAs in Cancer and Diseases
Fig 1. The workflow of eccDNA Sequencing
1. Genomic DNA extraction
2. Linearization of mitochondrial DNA
3. Removal of linear DNA
4. Rolling cycle amplification
5. Ultrasound fragmentation and DNA sequencing library construction
6. High-throughput sequencing
7. Data analysis and report
8. Delivered results
Annotated eccDNA data table
Histogram of eccDNA size distribution
Distribution of eccDNAs across the genome