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

  • Anna A. Neschastnova , Victoria K. Gasanova , Vladimir I. Popenko , Andreana Lambrinakos , Gennady A. Belitsky , Richard G.H. Cotton and Marianna G. Yakubovskaya
Published/Copyright: March 17, 2006
Biological Chemistry
From the journal Volume 387 Issue 3

Abstract

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 χ-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.

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Published Online: 2006-03-17
Published in Print: 2006-03-01

©2006 by Walter de Gruyter Berlin New York

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