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As a summary, a typical DNA rolling circle replication has five steps: [2] Circular dsDNA will be "nicked". The 3' end is elongated using "unnicked" DNA as leading strand (template); 5' end is displaced. Displaced DNA is a lagging strand and is made double stranded via a series of Okazaki fragments. Replication of both "unnicked" and displaced ...
The genome is not segmented and contains a single molecule of circular, negative-sense, single-stranded DNA.The complete genome is 3000–4000 nucleotides long. [1] They also contain a non-coding region with one to two 80–110 nt sequences that contain high GC content, forming a secondary structure of stems and loops. [2]
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 rolling circle mechanism that produces linear strands while progressing in a loop around the circular genome is also common. [6] [7] Some dsDNA viruses use a strand displacement method whereby one strand is synthesized from a template strand, and a complementary strand is then synthesized from the prior synthesized strand, forming a dsDNA ...
For imaging systems which cannot detect single fluorescence events, amplification of DNA templates is required. The three most common amplification methods are emulsion PCR (emPCR), rolling circle and solid-phase amplification. The final distribution of templates can be spatially random or on a grid.
[1] [2] [3] Diseases associated with this family include: bright yellow mosaic, yellow mosaic, yellow mottle, leaf curling, stunting, streaks, reduced yields. [ 2 ] [ 4 ] They have single-stranded circular DNA genomes encoding genes that diverge in both directions from a virion strand origin of replication (i.e. geminivirus genomes are ambisense ).
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The observed DNA replication intermediates included circular and branched circular concatemeric structures that likely arose by rolling circle replication. When assembling concatemers from synthetic oligonucleotides, increasing salt concentration to 200 mM was found to be a major optimizing factor due to its ability to enhance ionic strength ...