Analysis of DNA replication parameters in isolated organs of drosophila melanogaster by molecular combing

Ivankin, Anton V.; Колесникова, Т. Д.; Демаков, С. А.; Andreenkov, O. V.; Bildanova, E. R.; Andreenkova, N. G.; Жимулев, И. Ф.

doi:10.1134/s1990519x11010081

Cited by 1 publication

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“…In samples prepared using this method, it will be possible to identify the sites of DNA replication initiation through fluorescent in situ hybridization using specific probes for the genomic region of interest combined with the detection of the incorporated halogenated nucleotides. Previous studies have reported the use of molecular combing technology for the analysis of DNA replication in D. melanogaster (Kolesnikova et al, 2009(Kolesnikova et al, , 2010Ivankin et al, 2011), but in contrast to the method implanted in this study, those procedures involved prior isolation of nuclei from the analyzed tissues. B.…”

Section: Resultsmentioning

confidence: 67%

Molecular combing in the analysis of developmentally regulated amplified segments of Bradysia hygida

Passos

Togoro²,

Carignon³

et al. 2012

Genet. Mol. Res.

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ABSTRACT. Molecular combing technology is an important new tool for the functional and physical mapping of genome segments. It is designed to identify amplifications, microdeletions, and rearrangements in a DNA sequence and to study the process of DNA replication. This technique has recently been used to identify and analyze the dynamics of replication in amplified domains. In Bradysia hygida, multiple amplification initiation sites are predicted to exist upstream of the BhC4-1 gene. However, it has been impossible to identify them using the available standard techniques. The aim of this study was to optimize molecular combing technology to obtain DNA fibers from the polytene nuclei of the salivary glands of B. hygida to study the dynamics of DNA replication in this organism. Our results suggest that combing this DNA without prior purification of the polytene nuclei is possible. The density, integrity, and linearity of the DNA fibers were analyzed, fibers 50 to 300 kb in length were detected, and a 9-kb fragment within the amplified region was visualized using 2061 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 11 (3): 2060-2070(2012 Molecular combing analysis in Bradysia hygida biotin detected by Alexa Fluor 488-conjugated streptavidin technique. The feasibility of physically mapping these fibers demonstrated in this study suggests that molecular combing may be used to identify the replication origin of the BhC4-1 amplicon.

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Section: Resultsmentioning

confidence: 67%