Spontaneous DNA-DNA interaction of homologous duplexes and factors affecting the result of heteroduplex formation.
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 chi-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.
Institute of Carcinogenesis, Blokhin Cancer Research Centre, Russian Academy of Medical Sciences, Kashirskoye Shosse 24, Moscow 115478, Russia. myakubovskaya@crc.umos.ru
Anna A
AA
Neschastnova
Victoria K
VK
Gasanova
Vladimir I
VI
Popenko
Andreana
A
Lambrinakos
Gennady A
GA
Belitsky
Richard G H
RG
Cotton
Marianna G
MG
Yakubovskaya
D000818:Animals; D020137:Base Pair Mismatch / Q000235:genetics; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D004587:Electrophoresis, Agar Gel; D051379:Mice; D008854:Microscopy, Electron; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
D009692:Nucleic Acid Heteroduplexes; D004247:DNA