Molecular combing method in the research of DNA replication parameters in isolated organs of Drosophyla melanogaster
Ivankin, A.V.; Kolesnikova, T.D.; Demakov, S.A.; Andreenkov, O.V.; Bil'danova, E.R.; Andreenkova, N.G.; Zhimulev, I.F.
Tsitologiia 53(1): 75-82
2011
ISSN/ISBN: 0041-3771 PMID: 21473123 Document Number: 650465
Methods of physical DNA mapping and direct visualization of replication and transcription in specific regions of genome play crucial role in the researches of structural and functional organization of eukaryotic genomes. Since DNA strands in the cells are organized into high-fold structure and present as highly compacted chromosomes, the majority of these methods have lower resolution at chromosomal level. One of the approaches to enhance the resolution and mapping accuracy is the method of molecular combing. The method is based on the process of stretching and alignment of DNA molecules that are covalently attached with one of the ends to the cover glass surface. In this article we describe the major methodological steps of molecular combing and their adaptation for researches of DNA replication parameters in polyploidy and diploid tissues of Drosophyla larvae.