Design of DNA rolling-circle templates with controlled fork topology to study mechanisms of DNA replication.
Rolling-circle DNA amplification is a powerful tool employed in biotechnology to produce large from small amounts of DNA. This mode of DNA replication proceeds via a DNA topology that resembles a replication fork, thus also providing experimental access to the molecular mechanisms of DNA replication. However, conventional templates do not allow controlled access to multiple fork topologies, which is an important factor in mechanistic studies. Here we present the design and production of a rolling-circle substrate with a tunable length of both the gap and the overhang, and we show its application to the bacterial DNA-replication reaction.
['Monachino|Enrico|E|', 'Ghodke|Harshad|H|', 'Spinks|Richard R|RR|', 'Hoatson|Ben S|BS|', 'Jergic|Slobodan|S|', 'Xu|Zhi-Qiang|ZQ|', 'Dixon|Nicholas E|NE|', 'van Oijen|Antoine M|AM|']
D004261:DNA Replication D004269:DNA, Bacterial D004270:DNA, Circular D004926:Escherichia coli D021141:Nucleic Acid Amplification Techniques D009690:Nucleic Acid Conformation D013698:Templates, Genetic
DNA replication Nucleic-acid biochemistry Replisome Rolling-circle amplification