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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 ...
DNA molecules with a length of 1 kb are selected by loading on the 6% TBE PAGE gel. In the next step, the DNA molecules are circularized with T-tailed 30 bp long synthetic oligonucleotides (T30), which contains two outward-facing MmeI recognition sites, and the resulting circularized DNA undergoes rolling circle replication. The amplified ...
Workflow for DNA nanoball sequencing [1] DNA nanoball sequencing is a high throughput sequencing technology that is used to determine the entire genomic sequence of an organism. The method uses rolling circle replication to amplify small fragments of genomic DNA into DNA nanoballs. Fluorescent nucleotides bind to complementary nucleotides and ...
Prokaryotic DNA Replication is the process by which a prokaryote duplicates its DNA into another copy that is passed on to daughter cells. [1] Although it is often studied in the model organism E. coli, other bacteria show many similarities. [2] Replication is bi-directional and originates at a single origin of replication (OriC). [3]
[1] DNA sequence elements within oriC that are important for its function include DnaA boxes, a 9-mer repeat with a highly conserved consensus sequence 5' – TTATCCACA – 3', [2] that are recognized by the DnaA protein. DnaA protein plays a crucial role in the initiation of chromosomal DNA replication. [3]
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 ...
Salmonella virus P22 is a bacterial virus (bacteriophage) that infects Salmonella typhimurium. [1] Like many phages, it has been used in molecular biology to induce mutations in cultured bacteria and to introduce foreign genetic material. [2] P22 has been used in generalized transduction and is an important tool for investigating Salmonella ...
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 ...