How does rolling circle amplification work?

Rolling circle amplification (RCA) is an isothermal enzymatic process where a short DNA or RNA primer is amplified to form a long single stranded DNA or RNA using a circular DNA template and special DNA or RNA polymerases.

What is the rolling circle method?

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.

What are the different roles of phi29 DNA polymerase?

“Two positively charged residues of phi29 DNA polymerase, conserved in protein-primed DNA polymerases, are involved in stabilisation of the incoming nucleotide”. J Mol Biol. 335 (2): 481–94. doi:10.1016/j.

Is rolling circle DNA replication Semiconservative?

This result, which is indicative of semiconservative rolling circle replication of the circular plasmid, is the same as that previously observed with extracts of insect cells multiply infected with baculoviruses recombinant for the HSV-1-en- coded DNA polymerase, UL42 protein, helicase-primase, and ICP8 (8).

What is rolling circle model of DNA replication?

The rolling circle model for DNA replication suggests that DNA must be circular in order to be copied completely; the basic mode of reproducing an entire genome is to copy it from a circular template, using the circularity in an intrinsic way to guarantee that all of the genetic information is preserved.

What is the function of DNA polymerase III?

DNA Polymerase III, Bacterial The main function of the third polymerase, Pol III, is duplication of the chromosomal DNA, while other DNA polymerases are involved mostly in DNA repair and translesion DNA synthesis.

What viruses use rolling circle replication?

Hepatitis delta virus (HDV) contains a viroid-like circular RNA that is presumed to replicate via a rolling circle replication mechanism mediated by cellular RNA polymerases.

What is the function of Okazaki fragments?

Okazaki fragments are short sequences of DNA nucleotides (approximately 150 to 200 base pairs long in eukaryotes) which are synthesized discontinuously and later linked together by the enzyme DNA ligase to create the lagging strand during DNA replication.