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Anna A. Neschastnova, Victoria K. Gasanova, Vladimir I. Popenko, Andreana Lambrinakos, Gennady A. Belitsky, Richard G.H. Cotton, Marianna G. Yakubovskaya

Spontaneous DNA-DNA interaction of homologous duplexes and factors affecting the result of heteroduplex formation

Keywords: DNA branching, DNA-DNA interaction, heteroduplex formation, Holliday junctions

Mutation detection and mismatch repair investigations based on heteroduplex formation require a linear DNA structure. DNA branching, described previously under physiological conditions, has been analysed in the heteroduplex formation process. Symmetrical ?-structures were detected after heteroduplex formation by gel electrophoresis and electron microscopy. Buffer composition, DNA concentration and duplex end-sequences influence DNA branching. Duplexes with homologous central regions but non-complementary ends do not form hybrid heteroduplexes or hybrid Holliday junctions. Our results explain the requirements for efficient heteroduplex formation, which were previously determined empirically: special solution composition, optimal DNA concentration and GC clamps. This provides the theoretical background for further optimisation of the procedure.

Biological Chemistry, Walter de Gruyter

Print ISSN: 1431-6730
Volume: 387, 03/2006
Pages: 257 - 261

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