Extrachromosomal Circular DNA are composed of chromatin and replicate in the nucleus of the cell during cell division. Unlike typical chromosomes, they are composed of circular fragments of DNA, up to only a few million base pairs in size and contain no centromere or telomere. Extrachromosomal Circular DNA Extrachromosomal circular DNA (eccDNA) is circular DNA found in the nuclei of plant and animal cells, including human cells, in addition to chromosomal DNA. eccDNA can be from 200 up to 5 million base pairs in length. eccDNA has been found in the nuclei of human cancer cells and shown to carry many copies of
Single-cell DNA template strand sequencing cells are treated with bromodeoxyuridine (BrdU) which is then incorporated into their nascent DNA, acting as a substitute for thymidine. After at least one replication event has occurred, the daughter cells are synchronized at the G2 phase and individually separated by fluorescence-activated cell sorting (FACS). The cells are directly sorted into lysis buffer and their DNA is extracted. Having been arrested at a specified number of generations (usually one), the inheritance patterns of sister chromatids can be assessed. The following methods concentrate on the DNA sequencing of a single daughter cell’s DNA. At this point the chromosomes are composed of