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Rolling circle replication (RCR) is a process of unidirectional nucleic acid replication that can rapidly synthesize multiple copies of circular molecules of DNA or RNA, such as plasmids, the genomes of bacteriophages, and the circular RNA genome of viroids. Some eukaryotic viruses also replicate their DNA or RNA via the rolling circle mechanism.
Although the mechanism of replication has not been studied heavily, anelloviridae appears to use the rolling circle mechanism where first ssDNA is converted to dsDNA. It requires a host polymerase for replication to occur as the genome itself does not encode for a viral polymerase and, as a result, anelloviridae must replicate inside the cell's ...
DNA nanoballs are simply formed by denaturing double stranded, adapter ligated libraries and ligating the forward strand only to a splint oligonucleotide to form a ssDNA circle. Faithful copies of the circles containing the DNA insert are produced utilizing Rolling Circle Amplification that generates approximately 300–500 copies.
In the single stranded DNA viruses—a group that includes the circoviruses, the geminiviruses, the parvoviruses and others—and also the many phages and plasmids that use the rolling circle replication (RCR) mechanism, the RCR endonuclease creates a nick in the genome strand (single stranded viruses) or one of the DNA strands (plasmids).
A random hexamer or random hexonucleotides are for various PCR applications such as rolling circle amplification to prime the DNA.. They are oligonucleotide sequences of 6 bases which are synthesised entirely randomly to give a numerous range of sequences that have the potential to anneal at many random points on a DNA sequence and act as a primer to commence first strand cDNA synthesis.
A concatemer is a long continuous DNA molecule that contains multiple copies of the same DNA sequence linked in series. These polymeric molecules are usually copies of an entire genome linked end to end and separated by cos sites (a protein binding nucleotide sequence that occurs once in each copy of the genome).
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The circular fragments are copied by rolling circle replication resulting in many single-stranded copies of each fragment. The DNA copies concatenate head to tail in a long strand, and are compacted into a DNA nanoball. The nanoballs are then adsorbed onto a sequencing flow cell.
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